|
Fig. 5 TMEM47 degrades MAVS and STING via the autophagy-lysosome-dependent pathway. (A–C) Overexpression of the TMEM47 degrades MAVS and STING in a dose-dependent manner. EPC cells were seeded in six-well plates overnight and transfected with the indicated plasmids (1 µg each) (A) or plus TMEM47 (1 or 2 µg) (B and C) for 24 h. The cell lysates were subjected to IB with anti-Myc, anti-HA, and anti-β-actin Abs. (D and H) Effects of inhibitors on TMEM47-mediated degradation of MAVS and STING. EPC cells were seeded in six-well plates overnight and transfected with the indicated plasmids (1 µg each). At 18 h post-transfection, the cells were treated with dimethyl sulfoxide (DMSO), MG132 (20 µM), 3-MA (2 mM), Baf-A1 (100 nM), or CQ (100 µM) for 6 h. Then, the cells were harvested for IB with the Abs indicated. (E–G and I–K) TMEM47-induced MAVS/STING degradation is rescued by 3-MA, Baf-A1, and CQ in a dose-dependent manner. EPC cells were seeded in six-well plates overnight and co-transfected with the indicated plasmids. At 18 h post-transfection, the cells were treated with 3-MA (1 or 2 mM), Baf-A1 (50 or 100 nM), or CQ (50 or 100 µM) for 6 h. Then, the cells were harvested for IB with the indicated Abs. All experiments were repeated for at least three times with similar results.