Non-syndromic hearing impairment protein 5 is a protein that in humans is encoded by the DFNA5 gene. Hearing impairment is a heterogeneous condition with over 40 loci described. The protein encoded by this gene is expressed in fetal cochlea, however, its function is not known. Nonsyndromic hearing impairment is associated with a mutation in this gene. The observation that DFNA5 is epigenetically inactivated in a large number of cancers of frequent types (gastric, colorectal, and breast) is another important finding and is in line with its apoptosis-inducing properties. Indeed, if apoptosis is an intrinsic feature of DFNA5, shutting the gene down in tumor cells makes them more susceptible to uncontrolled cellular growth. Moreover, the fact that DFNA5 is regulated by P53 strongly suggests that DFNA5 is a tumor suppressor gene.
背景文献
1. Wei Y et al. GSDME-mediated pyroptosis promotes the progression and associated inflammation of atherosclerosis. Nat Commun. 2023 Feb
2. Ai YL et al. Mannose antagonizes GSDME-mediated pyroptosis through AMPK activated by metabolite GlcNAc-6P. Cell Res. 2023 Dec
Western blot analysis of DFNA5 / GSDME on different lysates with Rabbit anti-DFNA5 / GSDME antibody (HA723113) at 1/2,000 dilution.
Lane 1: SH-SY5Y cell lysate Lane 2: SH-SY5Y treated with 4μg/mL Doxorubicin for 24 hours cell lysate Lane 3: SW480 cell lysate (negative)
Lysates/proteins at 20 µg/Lane.
Predicted band size: 55 kDa Observed band size: 55/25 kDa
Exposure time: 3 minutes; 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 (HA723113) at 1/2,000 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.
☑ Cell treatment (CT)
Western blot analysis of DFNA5 / GSDME on different lysates with Rabbit anti-DFNA5 / GSDME antibody (HA723113) at 1/2,000 dilution.
Lane 1: 293T transfected with empty control cell lysate Lane 2: 293T transfected with mouse GSDME (C-terminal) cell lysate Lane 3: 293T transfected with mouse GSDME (N-terminal) cell lysate
Lysates/proteins at 10 µg/Lane.
Exposure time: 51 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 (HA723113) at 1/2,000 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.
☑ Cell treatment (CT)
DFNA5 / GSDME was immunoprecipitated from 0.2 mg SH-SY5Y treated with 4μg/mL Doxorubicin for 24 hours cell lysate with HA723113 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723113 at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: SH-SY5Y treated with 4μg/mL Doxorubicin for 24 hours cell lysate (input) Lane 2: HA723113 IP in SH-SY5Y treated with 4μg/mL Doxorubicin for 24 hours cell lysate Lane 3: Rabbit IgG instead of HA723113 in SH-SY5Y treated with 4μg/mL Doxorubicin for 24 hours cell lysate