The inhibition of mTOR causes the proteasomal degradation of Vif.

作者：Yuexuan Chen (11424022), Susanne Klute, Anju Bansal (56201), Konstantin M. J. Sparrer, Ruth Serra-Moreno · 发表于：Figshare · 年份：2026 · DOI：10.1371/journal.ppat.1014020.g002 · 研究领域：Chemistry、Molecular biology、Biology、Cell biology、Biochemistry

(A) HEK293T cells were transfected with 2,000 ng of an empty vector (V) or a NL4-3 Flag-Vif construct. 44 h later, cells were treated with Torin2 (5-10 nM) with or without BafA1 (500 nM) for 4 h. Next, cells were harvested, analyzed by western blot, and autophagy flux was measured by determining the LC3-II/I ratios and SQSTM1 levels relative to the vector-transfected and DMSO-treated cells. Data correspond to the mean and the SEM of 4 independent experiments. Blots are representative of 3 independent experiments. (B) Parental (wild type; WT) and ATG5 KO HEK293T cells were transfected with 2,000 ng of a NL4-3 Flag-Vif construct. 44 h later, cells were treated with DMSO or increasing concentrations of Rapamycin (1.3-6 μM) or Torin2 (2.5-10 nM) at two-fold increments. 4 h later, cells were harvested for their analysis by western blotting. Blots are representative of 3 independent experiments confirming ATG5 depletion and its effects on SQSTM1 levels, the lipidation of LC3 as well as on Flag-Vif. ACTB was included as a loading control. (C) The LC3-II/I ratios, the SQSTM1 and Vif levels across the Rapamycin treatment range relative to DMSO were determined by densitometry assays of western blots from 3 independent assays. SQSTM1 and Vif levels were normalized over ACTB prior to normalization to DMSO. Data represent the mean and the SEM of 3 independent experiments. Dotted lines represent 2-fold threshold. (D) The LC3-II/LC3-I ratios, the SQSTM1 and Vif levels across the T...