Transmembrane glycoprotein of the endoplasmic reticulum that functions as a transcription activator and initiates the unfolded protein response (UPR) during endoplasmic reticulum stress. Cleaved upon ER stress, the N-terminal processed cyclic AMP-dependent transcription factor ATF-6 alpha translocates to the nucleus where it activates transcription of genes involved in the UPR. Binds DNA on the 5'-CCAC[GA]-3'half of the ER stress response element (ERSE) (5'-CCAAT-N9-CCAC[GA]-3') and of ERSE II (5'-ATTGG-N-CCACG-3'). Binding to ERSE requires binding of NF-Y to ERSE. Could also be involved in activation of transcription by the serum response factor. May play a role in foveal development and cone function in the retina.
背景文献
1. Kohl S et al. Mutations in the unfolded protein response regulator ATF6 cause the cone dysfunction disorder achromatopsia. Nat Genet 47:757-765 (2015).
2. Lynch J M et al. A thrombospondin-dependent pathway for a protective ER stress response. Cell 149:1257-1268 (2012).
Western blot analysis of ATF6 on different lysates with Mouse anti-ATF6 antibody (HA601321) at 1/1,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: Jurkat cell lysate (20 µg/Lane) Lane 3: RAW264.7 cell lysate (20 µg/Lane) Lane 4: PC-12 cell lysate (20 µg/Lane) Lane 5: Mouse brain tissue lysate (40 µg/Lane) Lane 6: Mouse kidney tissue lysate (40 µg/Lane)
Predicted band size: 75 kDa Observed band size: 75 kDa
Exposure time: 1 minute 21 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601321) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of HeLa cells labeling ATF6 with Mouse anti-ATF6 antibody (HA601321) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-ATF6 antibody (HA601321) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
☑ Knockdown (KD)
Western blot analysis of ATF6 on different lysates with Mouse anti-ATF6 antibody (HA601321) at 1/2,000 dilution.
Lane 1: A549-WT cell lysate Lane 2: A549-KD ATF6 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 75 kDa Observed band size: 75 kDa
Exposure time: 11 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601321) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
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