Fig. 2. mTOR signaling regulates polκ subcellular localization.
(A) Immunofluorescence staining of A375 cells treated with DMSO or 5 μM PLX4032 for 24h hours. Scale bar, 10 μm. (B) Quantification of the percentage of cells with phosphorylated S6 (p-S6, S240/244). Mean ± S.E.M; at least 400 cells were counted for each sample across 8 fields of view. ***P < 0.001, paired two-tailed t-test. (C) Immunofluorescence staining of A375 cells treated with 5 μM PLX4032 for 3, 6, 12, or 24 hours. The bar graph shows the percentage of cells with nuclear enrichment of polκ (left axis, solid line) and the percentage of cells with phosphorylated S6 (p-S6, right axis, dashed line), Mean ± S.E.M; at least 150 cells were counted for each sample across 4–8 fields of view. The Pearson correlation coefficient (r) was calculated for the percentage of cells with nuclear enrichment of polκ vs. the percentage of cells with phosphorylated S6. (D) Immunofluorescence staining of A375 cells treated with DMSO, 0.5 μM rapamycin, 5 μM PP242, or 30 μM LY294002 for 6 hours. Scale bar, 10 μm. (E) Quantification of the percentage of cells with nuclear enrichment of polκ after 6 hours of treatment. Mean ± S.E.M; at least 200 cells were counted for each sample across 8–12 fields of view. **P < 0.01, ***P < 0.001, paired two-tailed t-test.