Journal: PLOS Pathogens

Article Title: HIV-1 Vpr is targeted for degradation by autophagy

doi: 10.1371/journal.ppat.1014020

Figure Lengend Snippet: HEK293T cells were transfected with 2,000 ng of either Lab-Vpr or TFV-Vpr. 48 h later, cells were harvested, and lysates were immunoprecipitated for BNIP3L (A) , OPNT (B) , NBR1 (C) , SQSTM1 (D) , TAX1BP1 (E) , or NDP52 (F) . The pulldown fraction was analyzed for each of these autophagy receptors and Vpr. The whole cell lysates were also analyzed for these autophagy factors, Vpr and ACTB. Blots are representative of 3 independent experiments. (G) Lab-Vpr was transfected in parental, SQSTM1 KO / TAX1BP1 KO and SQSTM1 KO / TAX1BP1 KO / NDP52 KO knockout cells. 44 h later, cells were treated with increasing amounts of Torin2 (5-10 nM) for 4 h. Cells were then harvested and analyzed by western blot for Vpr, SQSTM1, TAX1BP1, NDP52 and ACTB. Blots are representative of 3 independent experiments. Graph: Vpr levels were quantified by normalizing to ACTB, and their relative expression compared to WT cells treated with DMSO (D on the x axis) were calculated from 3 independent experiments. Data correspond to the mean and the SEM of 3 independent experiments. *: p < 0.05.

Article Snippet: NBR1 , Rabbit monoclonal (E6Q3F) to NBR1 , 1:1000 , Cell signaling, #20145S.

Techniques: Transfection, Immunoprecipitation, Knock-Out, Western Blot, Expressing