FIG 6.

The ARV p17 protein inhibits mTORC1 translocation to lysosomes. (A) To study whether p17, PRAS40, TSC2, and insulin regulate mTORC1 translocation to lysosomes, Vero cells were fixed and processed for immunofluorescence staining with DAPI. Vero cells were pretreated with insulin (0.2 μM) for 2 h, followed by transfection with the pcDNA3.1-p17 vector for 24 h. All transfections or cotransfections were performed for 24 h. Stained mTOR (red) and lysosome (green) were observed under a fluorescence microscope. The yellow dots in the merged panel show that the mTOR complex is located on the lysosome. (B) Quantification of colocalization was quantified with ImageJ software. The amount of colocalized fluorescence in the mock-treated control group was considered to be 1-fold. At least 10 fields of 2 cells per field were taken for each sample per experiment. All experiments were performed in three independent experiments.