Figure 6. FNIP knockdown results in increased mTOR lysosomal localization and diffusion of FLCN in HeLa cells.
A. Immunoblot (IB) analysis of whole cell lysates derived from HeLa cells stably expressing shRNA against GFP (control), FNIP1, FNIP2, or FNIP1/FNIP2. After being serum and amino acid-starved for 3 h, cells were treated with fresh 10% FBS DMEM and harvested at the indicated time points. B-E. Confocal images of HeLa cells presented in (A). DAPI-loaded HeLa cells were analyzed for co-localization of FLCN (B and D) (red) or mTOR (C and E) (red) with a lysosomal marker, LAMP1 (green). Y (yellow) indicates predominant localization of FLCN or mTOR in the lysosome. Scale bars, 20 μm (5 μm in the enlarged images).