Supplementary Figure 7: Nuclear accumulation of RPS14 in senescence.

(a-c) Indirect immunofluorescence (IF) with specific anti-RPS14 antibody and nuclear counterstaining with DAPI. Images from (a) IMR90 cells expressing H-RASV12 (RAS) (1 out of 3 independent experiments with similar results), PML-IV (PML) (1 out of 2 independent experiments with similar results) or an empty control vector at day 12 post-infection, (b) young (passage 25) and old (passage 43) IMR90 cells (1 out of 2 independent experiments with similar results) and (c) IMR90 cells treated with DMSO (CTRL) or camptothecin in DMSO (final concentration in medium of 35 nM) for 3 days (1 out of 2 independent experiments with similar results). Data in RPS14 panel were quantified from 100 cell counts in triplicate and are presented as the mean percentage of cells with RPS14 nuclear staining. (d) Scale for intensity of staining by indirect IF with specific anti-RPS14 as in (a-c). (e-g) Hyperresolution structure illumination microscopy of immunofluorescence staining using a specific anti-RPS14 antibody and counterstaining with DAPI. (e) Comparison of IMR90 cells expressing H-RASV12 (RAS) or an empty control vector and fixed at day 12 post-infection. CTRL: 2 cells out of 15 showed nuclear staining; RAS: 5 cells out of 6 showed nuclear staining. (f) Comparison of IMR90 cells treated with DMSO (CTRL) or camptothecin in DMSO (final concentration in medium of 35 nM) for 3 days. DMSO: 2 cells out of 24 showed nuclear staining; Camptothecin: 4 cells out of 5 showed nuclear staining. (g) Comparison of young (passage 25) or old (passage 43) IMR90 cells. Young: 1 cell out of 10 showed nuclear staining; Old: 4 cells out of 4 showed nuclear staining.