Endoplasmic reticulum (ER)-anchored autophagy regulator which mediates ER delivery into lysosomes through sequestration into autophagosomes . Promotes membrane remodeling and ER scission via its membrane bending capacity and targets the fragments into autophagosomes via interaction with ATG8 family proteins . Active under basal conditions. Required for collagen quality control in a LIR motif-dependent manner (By similarity). Required for long-term survival of nociceptive and autonomic ganglion neurons.
背景文献
1. Kohno S. et al.2019. An N-terminal-truncated isoform of FAM134B (FAM134B-2) regulates starvation-induced hepatic selective ER-phagy. Life Sci Alliance. 17;2(3):e201900340.
2. Mookherjee D. et al. 2021. RETREG1/FAM134B mediated autophagosomal degradation of AMFR/GP78 and OPA1 -a dual organellar turnover mechanism. Autophagy. 17(7):1729-1752.
Western blot analysis of FAM134B on different lysates with Rabbit anti-FAM134B antibody (HA721752) at 1/1,000 dilution.
Lane 1: HEK-293 cell lysate (30 µg/Lane) Lane 2: U-2 OS cell lysate (30 µg/Lane) Lane 3: MCF7 cell lysate (30 µg/Lane) Lane 4: HeLa cell lysate (30 µg/Lane) Lane 5: HepG2 cell lysate (30 µg/Lane) Lane 6: TT cell lysate (30 µg/Lane) Lane 7: Jurkat cell lysate (30 µg/Lane) Lane 8: NIH/3T3 cell lysate (30 µg/Lane) Lane 9: C2C12 cell lysate (30 µg/Lane) Lane 10: PC-12 cell lysate (30 µg/Lane) Lane 11: C6 cell lysate (30 µg/Lane) Lane 12: Mouse liver tissue lysate (30 µg/Lane) Lane 13: Rat liver tissue lysate (30 µg/Lane)
Predicted band size: 55 kDa Observed band size: 70 kDa
Exposure time: 6 minutes 20 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 (HA721752) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of Jurkat cells labeling FAM134B with Rabbit anti-FAM134B antibody (HA721752) at 1/100 dilution.
Cells were fixed in 80% precooled methanol 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 Rabbit anti-FAM134B antibody (HA721752) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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