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. 2016 Dec 25;8(6):9947–9960. doi: 10.18632/oncotarget.14221

Figure 5. Ectopic expression of non-degradable FNIP2 enhances FLCN lysosomal localization and diffusion of mTOR in HeLa cells.

Figure 5

A. Immunoblot (IB) analysis of whole cell lysates derived from HeLa cells transfected with empty vector (EV) or FNIP2-expression plasmid (WT or 3A: S237/242/244A). At 12 h post-transfection, cells were treated with 0.4 mg/mL G418 for 72 h to eliminate non-transfected cells. 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).