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Brefeldin A (BFA)

Synonyms: Cyanein, Decumbin

Brefeldin A (BFA) is a lactone antibiotic and ATPase inhibitor for protein transport with IC50 of 0.2 μM in HCT 116 cells, induces cancer cell differentiation and apoptosis. It could also improve the HDR(homology-directed repair) efficiency and be an enhancer of CRISPR-mediated HDR. Brefeldin A is also an inhibitor of autophagy and mitophagy.

Brefeldin A (BFA) Chemical Structure

Brefeldin A (BFA) Chemical Structure

CAS No. 20350-15-6

Purity & Quality Control

Brefeldin A (BFA) Related Products

Signaling Pathway

ATPase Signaling Pathways

ATPase Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
PC12 Function Assay 2 μM 1 h inhibits the L-DOPA (20 μM)-induced transient ERK1/2 phosphorylation  26363191
C2C12 Function Assay 1 μg/ml 1 h abolishes cytokine release from C2C12 myotubes 26291279
MEFs WT Function Assay 5 μM 20 min causes resident enzymes such as NAGT-GFP, to diffuse back to the ER 26196023
MEFs VAMP7 KO Function Assay 5 μM 20 min causes resident enzymes such as NAGT-GFP, to diffuse back to the ER 26196023
SMCs Function Assay 10 µg/ml 0-12 h DMSO shows a trend towards a higher concentration of the ER/SR network in the perinuclear area  26172080
SMCs Function Assay 10 µg/ml 0-12 h DMSO causes a transient Ca2+ release from the ER/SR  26172080
HEMC-1 Function Assay 0.1 µg/ml 24 h causes a higher inhibitory effect on exocytosis than nocodazole 25972759
HUVEC Function Assay 10 μM 1 h DMSO abolishes hypoxia-induced release of ATP from apical and basolateral surfaces 25956988
HUVEC Function Assay 10 μM 1 h DMSO increases the number and intensity of fluorescent areas especially in perinuclear space 25956988
Caco-2 Function Assay 2.5 μM 30 min attenuates the TGF-β1-mediated increase in SERT function 25954931
NRK Function Assay 200 ng/ml 4 h DMSO rescues mitotic progression 25948586
HeLa Function Assay 200 ng/ml 3 h DMSO induces the artificial break-up of the Golgi complex 25948586
COS Function Assay 1 μg/ml 3 h completely disperses the AP-1 signal  25915900
DF1  Function Assay 1 μM  48 h DMSO disperses the exogenous CSGalNAcT2 protein 25807054
nHDFs  Function Assay 1 μM  2 h prevents the assembly of cytosolic coat proteins onto Golgi membranes 25772616
FRT  Function Assay 5 μg/ml 2 h blocks trafficking through the Golgi complex by inhibiting ER-to-Golgi transport 25767115
FRT  Function Assay 5 μg/ml 2 h prevents the increase in cleaved α subunits when [Na+]i was reduced 25767115
HepG2  Function Assay 1 µM  24 h DMSO decreases the level of PXR mRNA 25616597
SMCs Function Assay 1μg/mL 3 h accumulates CNPY2 protein in the ER compartment and no longer co-localized with the Golgi marker  25589425
OB-6 Apoptosis Assay 2.7 μM 48 h induces apoptosis 25532480
iPSC-CMs  Function Assay 500 ng/ml 48 h increases the intensity of the higher mobility LAMPs at the cost of the lower mobility species 25488666
SP-Nluc Function Assay 5 mg/mL 6 h DMSO  causes an increase in reporter activity in the parasite 25392998
PEXEL-Nluc Function Assay 5 mg/mL 6 h DMSO  causes an increase in reporter activity in the parasite 25392998
H1299 Function Assay 10 μg/ml 24 h induces autophagy  25388970
MDA-MB-231 Cell Viability Assay 0–50 μg/mL 48 h EC50 = 0.016 µg/mL 25356567
MDA-MB-231 Apoptosis Assay 0.1 μg/mL 4 h induces apoptosis 25356567
MDA-MB-231 Growth Inhibition Assay 0.01/0.05 μg/mL 24 h increases the fraction of sub-G1 cell debris 25356567
MDA-MB-231 Apoptosis Assay 0.05–1 μg/mL 24 h induces PARP (poly ADP-ribose polymerase-1) cleavage 25356567
MDA-MB-231 Function Assay 0–50 μg/mL 24 h inhibits the formation of 3D and 2D colonies 25356567
A172 Function Assay 10 μg/ml 4 h DMSO  results in the retrograde transport of fluorescent granules 25239507
KMS-6 Function Assay 1 μM  24 h exhibits half the secretion of galanin-LI as did the control 25229126
MEC Function Assay 1 μM 1.5 h causes a dramatic decrease in the surface VEGFR2 25228815
HEK293/hERG Function Assay 10 μM 1 h results in a time-dependent reduction mature hERG protein  25218469
RBE4 Apoptosis Assay 2 μM 3–24 h induces apoptosis time dependently 25128025
RBE4 Function Assay 2 μM 3–24 h increases the XBP1 protein levels after 3 and 6 h of treatment 25128025
RBE4 Function Assay 2 μM 3–24 h increases active caspase-12 in a time-dependent manner  25128025
RBE4 Function Assay 2 μM 3–24 h increases the levels of ROS time-dependently 25128025
RBE4 Function Assay 2 μM 3–24 h induces a delayed depletion of the ER Ca2+ content at 6 h of incubation significantly 25128025
RBE4 Function Assay 2 μM 3–24 h induces an overload of Ca2+ in the mitochondria in the first 6 h of incubation (p < 0.001) but Ca2+ levels in this organelle decreased after 12 h of incubation 25128025
Huh-7  Function Assay 1μg/mL 3–24 h increases the level of APE1 in a time-dependent manner 25026174
HepG2  Function Assay 1μg/mL 3–24 h increases the level of APE1 in a time-dependent manner 25026174
H838-LKB1 Function Assay 30 ng/ml 12/18 h increases the protein levels of BiP  25011082
H838-KDLKB1  Function Assay 30 ng/ml 12/18 h increases the protein levels of BiP  25011082
H838-KDLKB1  Function Assay 30 ng/ml 12/18 h increases the levels of phosphorylated eIF2α (phospho-eIF2α) 25011082
3T3-L1 Function Assay 5 μg/ml 30 min mimics the effects of insulin and causes robust phosphorylation of Akt (Ser 473) and phosphorylation of AS160 (Thr 642 and Ser 588) 24843827
3T3-L1 Function Assay 5 μg/ml 30 min recapitulates insulin action with respect to regulating Akt activity and AS160 phosphorylation 24843827
3T3-L1 Function Assay 5 μg/ml 30 min causes reversible redistribution of GLUT4 24843827
3T3-L1 Function Assay 5 μg/ml 1 h causes redistribution of GLUT4 but not increase in glucose uptake 24843827
3T3-L1 Function Assay 5 μg/ml 1 h causes phosphorylation of the FoxO1 transcription factor 24843827
HeLa  Function Assay 5 μg/ml 3 h causes nuclear exclusion of the FoxO1 transcription factor and decreases transcription of FoxO1-regulated genes 24843827
HEK293 Function Assay 5 μg/ml 12 h abolishes CMA-induced CRELD2 secretion 24687431
COS-1 Function Assay 5 µg/ml 24 h  restricts localization of NB in the perinuclear region  24671751
PRP Function Assay 10 μM abrogates SDF-1α-mediated CXCR7 externalization  24668750
RAW264.7 Apoptosis Assay 4 μM 48 h attenuates the inhibition of ox-LDL-induced apoptosis and the facilitation of cholesterol efflux by Ac-hE-18A-NH2 24639032
MDMs Apoptosis Assay 10 μg/ml 12/15 h induces apoptosis 24556695
PMHs  Function Assay 10–20 μg/ml 24 h DMSO induced ER stress 24407242
PMHs  Apoptosis Assay 10–20 μg/ml 24 h DMSO increases cell death 24407242
HEK293/tau Function Assay 5 μM 1/2/4 h induces Golgi fragmentation  24368089
HEK293/tau Function Assay 5 μM 3 h induces tau hyperphosphorylation 24368089
ADF Function Assay 10 μM 16 h inhibits the ZnCl2-induced translocation of CRT 24228232
U373  Function Assay 10 μM 16 h inhibits the ZnCl2-induced translocation of CRT 24228232
RKO-HIPK2i Function Assay 10 μM 16 h inhibits the ZnCl2-induced translocation of CRT 24228232
ADF  Function Assay 10 μM  6 h impairs the DC activation 24228232
Huh7 Function Assay 5 μg/ml 4 h abolishes the secretion of intracellular ApoB 24100140
Huh7 Function Assay 5 μg/ml 1 h causes a significant increase in ApoB-crescents 24100140
Huh7 Function Assay 5–10 ng/ml 12 h increases ApoB-crescents without inhibiting secretion 24100140
BAECs Function Assay 5 μg/ml 0-4 h induces the rapid dephosphorylation of eNOS at Ser1179 24085225
Macrophages Function Assay 71 µM 6 h inhibits lunasin internalization  24039740
Colo 205 Growth Inhibition Assay 0-5 μg/mL 48 h inhibits cell growth in suspension cultures with an estimated IC50 of ~15 ng/mL 23973996
Colo 205 Function Assay 0.012-0.025 μg/mL 14 d reduces the clonogenicity of Colo 205 CSCs 23973996
Colo 205 Apoptosis Assay 0.1 μg/mL 0-24 h induces apoptosis of Colo 205 cells in suspension cultures 23973996
Colo 205 Function Assay 0.015 μg/mL 24 h induces the expression of ER stress-related genes 23973996
Colo 205 Function Assay 0.015 μg/mL 24 h inhibits the activity of MMPs 23973996
IBRS2 Function Assay 5 μg/ml 0.5 h DMSO disrupts the ERGIC and Golgi  23963534
IBRS2 Function Assay 5 μg/ml 0.5 h DMSO enhances FMDV infection 23963534
HeLa Function Assay 2 μM 2 h  attenuates the TNF-induced secretion of IL-15 23950892
HFS  Function Assay 0-1 μg/ml 24 h GLTP expression reaches a plateau at concentrations as low as 0.01 µg/ml 23894633
HFS  Function Assay 0.01 µg/ml 24 h increases the expression of glycosphingolipid synthase genes at 6 h 23894633
OVCAR-3 Growth Inhibition Assay 1–15 μM  24 h induces a loss of cell viability dose dependently  23826964
OVCAR-3 Function Assay 1–15 μM  24 h induces nuclear damage 23826964
OVCAR-3 Apoptosis Assay 1-10 μM 4 h induces the activation of apoptosis-related proteins 23826964
OVCAR-3 Apoptosis Assay 10 μM 24 h induces activation of caspases 23826964
OVCAR-3 Function Assay 1–10 μM 24 h induces disruption of the mitochondrial transmembrane potential 23826964
OVCAR-3 Function Assay 1–10 μM 24 h induces formation of reactive oxygen species 23826964
OVCAR-3 Function Assay 1–10 μM 24 h inhibits cell adhesion and migration 23826964
MKN45 Growth Inhibition Assay IC50<0.001 μg/ml 23793342
LOVO Growth Inhibition Assay IC50=0.12 μg/ml 23793342
A549 Growth Inhibition Assay IC50=0.04 μg/ml 23793342
MDA-MB-435 Growth Inhibition Assay IC50<0.001 μg/ml 23793342
HepG2 Growth Inhibition Assay IC50<0.001 μg/ml 23793342
HL-60 Growth Inhibition Assay IC50<0.001 μg/ml 23793342
neural precursor cells Function assay Inhibition of neurosphere proliferation of mouse neural precursor cells by MTT assay 17417631
HeLa Function assay 100 uM 2 hrs Dispersion of cis golgi marker betaCoP in human HeLa cells at 100 uM for 2 hrs 17563369
HeLa Function assay 100 uM 2 hrs Dispersion of cis golgi marker KDEL in human HeLa cells at 100 uM for 2 hrs 17563369
Vero Function assay 10 ug/ml 5 mins Inhibition of Arf1 in african green monkey Vero cells assessed as rapid AP-1 dispersal from golgi membranes at 10 ug/ml after 5 mins by immunofluorescence method 19182783
Vero Function assay 10 ug/ml 5 mins Inhibition of Arf1 in african green monkey Vero cells assessed as rapid GGA3 dispersal from trans golgi network at 10 ug/ml after 5 mins by immunofluorescence method 19182783
Vero Function assay 10 uM 1 hr Inhibition of GBF1 QNV deleted mutant in african green monkey Vero cells assessed as effect on change in golgi morphology at 10 uM after 1 hr by immunofluorescence method 19182783
Vero Function assay 10 uM 1 hr Inhibition of GBF1 QNV to AAA mutant in african green monkey Vero cells assessed as effect on change in golgi morphology at 10 uM after 1 hr by immunofluorescence method 19182783
Vero Function assay 10 ug/ml 1 hr Inhibition of Arf1 in african green monkey Vero cells assessed as decrease in Arf1-GTP levels at 10 ug/ml after 1 hr 19182783
Vero Function assay 10 uM Inhibition of GBF1 in african green monkey Vero cells assessed as inhibition of StxB-SS retrogade transport from endosomes to TGN at 10 uM by immunofluorescence method 19182783
Vero Function assay 10 uM 1 hr Inhibition of GBF1 in african green monkey Vero cells assessed as punctate and diffuse distribution of medial-Golgi marker giantin from TGN at 10 uM after 1 hr by immunofluorescence method 19182783
Vero Function assay 10 ug/ml 1 hr Inhibition of Arf1 in african green monkey Vero cells assessed as punctate and diffuse distribution of medial-Golgi marker giantin at 10 ug/ml after 1 hr by immunofluorescence method 19182783
Vero Function assay 10 ug/ml 5 mins Inhibition of Arf1 in african green monkey Vero cells assessed as rapid COPI redistribution from golgi at 10 ug/ml after 5 mins by immunofluorescence method 19182783
Vero Function assay 10 ug/ml 1 hr Inhibition of Arf1 in african green monkey Vero cells assessed as tubule formation from trans golgi network and endosomes before its dispersal at 10 ug/ml after 1 hr by immunofluorescence method 19182783
Vero Function assay 10 ug/ml 1 hr Inhibition of Arf1 in african green monkey Vero cells assessed as giantin positive punctate structures in contact with Sec31-positive ER exit site at 10 ug/ml after 1 hr by immunofluorescence method 19182783
Vero Function assay 10 ug/ml Inhibition of Arf1 in african green monkey Vero cells assessed as inhibition of StxB-SS retrogade transport from endosomes to TGN at 10 ug/ml by immunofluorescence method 19182783
Vero Function assay 10 uM 1 hr Induction of GBF1 in african green monkey Vero cells assessed as punctate and diffuse distribution of cis-Golgi marker GM130 from TGN at 10 uM after 1 hr by immunofluorescence method 19182783
Vero Function assay 10 ug/ml 1 hr Inhibition of Arf1 in african green monkey Vero cells assessed as punctate and diffuse distribution of cis-Golgi marker GM130 at 10 ug/ml after 1 hr by immunofluorescence method 19182783
NRK Function assay 7 uM 60 mins Golgi-disturbing activity in golgi apparatus of rat NRK cells assessed as fusion of golgi membrane fusion with endoplasmic reticulum at 7 uM after 60 mins by Hoechst 3342 staining-based immunofluorescence microscopy 20189813
NCI60 Cytostatic assay Cytostatic activity against human NCI60 cells by SRB assay, GI50=0.0206μM. 23805957
NCI60 Cytostatic assay Cytostatic activity against human NCI60 cells by SRB assay, TGI=3.48μM. 23805957
HeLa R19 Antiviral assay 0.5 uM 7 hrs Antiviral activity against Coxsackievirus B3 infected in human HeLa R19 cells assessed as inhibition of viral replication at 0.5 uM after 7 hrs by luciferase reporter gene assay 23805957
HeLa Function assay 5 uM 30 to 60 mins Induction of golgi apparatus disassembly in human HeLa cells at 5 uM after 30 to 60 mins by confocal microscopic analysis 23805957
Arabidopsis thaliana root cells Function assay 90 uM 30 mins Induction of morphological changes of golgi apparatus in Arabidopsis thaliana root cells expressing ST-YFP/VHAa1-RFP at 90 uM after 30 mins by confocal laser scanning microscopic analysis 23805957
HeLa R19 Antiviral assay 5 to 50 uM 7 hrs Antiviral activity against Coxsackievirus B3 infected in human HeLa R19 cells assessed as inhibition of viral replication at 5 to 50 uM after 7 hrs by luciferase reporter gene assay 23805957
PC3 Function assay 50 nM 72 hrs Potentiation of 3 nM docetaxel-induced cytotoxicity against human PC3 cells assessed as decrease in cell viability at 50 nM after 72 hrs by trypan blue exclusion assay 28462831
L02 Cytotoxicity assay 72 hrs Cytotoxicity against human L02 cells assessed as reduction in cell viability after 72 hrs by MTT assay, IC50<0.0004μM. 28494251
PC3 Antiproliferative assay 72 hrs Antiproliferative activity against human PC3 cells after 72 hrs by MTT assay, IC50=0.068μM. 28494251
HT-29 Antiproliferative assay 72 hrs Antiproliferative activity against human HT-29 cells after 72 hrs by MTT assay, IC50=0.16μM. 28494251
HepG2 Antiproliferative assay 72 hrs Antiproliferative activity against human HepG2 cells after 72 hrs by MTT assay, IC50=0.35μM. 28494251
LO2 Antiproliferative assay 72 hrs Antiproliferative activity against human LO2 cells after 72 hrs by MTT assay, IC50<0.001μM. 29524728
Bel7402 Antiproliferative assay 72 hrs Antiproliferative activity against human Bel7402 cells after 72 hrs by MTT assay, IC50=0.024μM. 29524728
HL60 Antiproliferative assay 72 hrs Antiproliferative activity against human HL60 cells after 72 hrs by MTT assay, IC50=0.025μM. 29524728
PC3 Antiproliferative assay 72 hrs Antiproliferative activity against human PC3 cells after 72 hrs by MTT assay, IC50=0.068μM. 29524728
Bel7402/5-FU Antiproliferative assay 72 hrs Antiproliferative activity against human Bel7402/5-FU cells after 72 hrs by MTT assay, IC50=0.82μM. 29524728
HeLa Function assay 18 uM 3 hrs Inhibition of alkaline phosphatase secretion in human HeLa cells at 18 uM incubated for 3 hrs 31421965
VERO-E6 Function assay 48 hrs Determination of IC50 values for inhibition of SARS-CoV-2 induced cytotoxicity of VERO-E6 cells after 48 hours exposure to 0.01 MOI SARS CoV-2 virus by high content imaging, IC50=0.02μM. ChEMBL
VERO-E6 Function assay 48 hrs Toxicity CC50 against VERO-E6 cells determined at 48 hours by high content imaging (same conditions as 2_LEY without exposure to 0.01 MOI SARS CoV-2 virus), CC50=0.06μM. ChEMBL
Click to View More Cell Line Experimental Data

Biological Activity

Description Brefeldin A (BFA) is a lactone antibiotic and ATPase inhibitor for protein transport with IC50 of 0.2 μM in HCT 116 cells, induces cancer cell differentiation and apoptosis. It could also improve the HDR(homology-directed repair) efficiency and be an enhancer of CRISPR-mediated HDR. Brefeldin A is also an inhibitor of autophagy and mitophagy.
Targets
ATPase (HCT 116) [1]
0.2 μM
In vitro
In vitro

Brefeldin A is a fungal metabolite and blocks the forward transport between the endoplasmic reticulum and Golgi apparatus, Brefeldin A causes an impaired distribution of the membrane proteins. When HCT 116 human colon cancer cell is treated with Brefeldin A, morphological changes indicating cell differentiation are observed. Brefeldin A exerts its cytotoxic effects mainly by inducing differentiation and apoptosis in tumor cells. [1]

The treatment of the strips with 20 μg/mL Brefeldin A for 6 hours completely abolishes the relaxation induced by bradykinin in the presence of 10mM indomethacin and 30 μM L-NOARG. The treatment with 20 μg/mL Brefeldin A substantially abolishes the bradykinin-induced decreases in [Ca2+]i and tension in the range of concentrations between 1 nM and 1 mM. Brefeldin A has no effect on the [Ca2+]i elevation in endothelial cells induced by bradykinin or substance P. [2]

Addition of the fungal metabolite Brefeldin A does not affect the spontaneous phospholipid-dependent GTPS binding to myr-rARF1 but totally abolishs the retinal isotonic extract (RIE)-catalyzed exchange, with half-maximal inhibition at 2 μM Brefeldin A. Brefeldin A prevents a wide variety of membrane traffic pathways. Brefeldin A inhibits an ADP-ribosylation factor-specific guanine nucleotide exchange activity present in Golgi membranes or in brain cytosol. The complete prevention by Brefeldin A strongly suggests that the retinal extract contains an ARF-specific guanine nucleotide exchange factor. Retinal isotonic extract (RIE)-catalyzed GTPS release from both ADP-ribosylation factors (ARFs) is only partly inhibited by Brefeldin A, even at 300 μM. [3]

Brefeldin A induces fusion of the Golgi apparatus with the ER. Brefeldin A abolishes the inhibitory effect of the CERT inhibitor HPA-12. Brefeldin A treatment, which induces fusion of the Golgi apparatus and the ER, rescues the limonoid-induced prevention of sphingomyelin biosynthesis. BFA treatment of CHO cells causes a 2 to 3 fold increase in sphingomyelin synthesis. [4]

Apart from B-CLL cells, Brefeldin A reportedly causes apoptosis in multiple myeloma (U266, NCI-H929), Jurkat, HeLa, leukaemia (HL60, K562, BJAB), colon (HT-29) and prostate, as well as adenoid cystic sarcoma cells. The administration of 25 ng/mL of Brefeldin A completely blocks growth of HF4.9 and HF28RA cells, whereas higher Brefeldin A doses (75 ng/mL) are required to achieve the same effect in HF1A3 cells. Cell proliferation is inhibited within 24 hours in a dose-dependent manner and, depending on the cell line, almost complete cessation of 3H-thymdine incorporation is observed at 50-75 ng/mL of Brefeldin A (26%, 76%, 87% inhibition at 50 ng/ml and 75%, 87%, 92% inhibition at 75 ng/mL for HF1A3, HF4.9 and HF28RA cells respectively. Brefeldin A-induced cell killing is in a dose-dependent manner using YO-PRO 1/PI assay. [5]

Brefeldin A could improve the HDR(homology-directed repair) efficiency. It is an enhancer of CRISPR-mediated HDR[6].
Cell Research Cell lines Human follicular lymphoma cell lines HF1A3, HF4.9 and HF28RA
Concentrations 0 ng/mL -75 ng/mL
Incubation Time 5 days
Method

HF1A3, HF4.9 cell viability upon the treatments is tested using double staining of cells with YO-PRO 1/PI and SYTO16/PI probes. To access cell proliferation, cells are treated with 0–100 ng/mL Brefeldin A in complete medium for 20 hours before adding 1 μCi/mL [methyl-3H]-thymidine for additional 4 hours at 37 °C. The incorporated radioactive thymidine is quantified by scintillation counting with Microbeta counter. To examine long-term effects of Brefeldin A treatment, cells are seeded at initial concentration 105 cells/mL and treated with 0-75 ng/mL Brefeldin A for up to 5 days. At the time indicated, a sample of cells is removed and viable cell number is assessed by standard Trypan Blue exclusion assay.
Experimental Result Images Methods Biomarkers Images PMID
Western blot p53 / GRP78 22859938
Immunofluorescence FMNL1 / GM130 ErbB3 / Calnexin MTP / GBF1 21868368
Growth inhibition assay Cell viability 28462831
In Vivo
In vivo

Brefeldin A (BFA) is a lactone antibiotic and a specific inhibitor of protein trafficking. Brefeldin A blocks the transport of secreted and membrane proteins from endoplasmic reticulum to Golgi apparatus.
Animal Research Animal Models C57BL/6 mice
Dosages 250 µg
Administration i.p.

Chemical Information & Solubility

Molecular Weight 280.36 Formula

C16H24O4
CAS No. 20350-15-6 SDF Download Brefeldin A (BFA) SDF
Smiles CC1CCCC=CC2CC(CC2C(C=CC(=O)O1)O)O
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 56 mg/mL ( (199.74 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble


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