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

Figure 4. β-TRCP knockdown leads to FNIP stabilization, increased FLCN lysosomal localization, and diffused mTOR distribution in HeLa cells.

Figure 4

A. Immunoblot (IB) analysis of whole cell lysates derived from HeLa cells stably expressing shRNA against GFP (control) or β-TRCP1+2 (shRNA against both β-TRCP1 and β-TRCP2). 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).