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. 2023 Nov 24;69:102972. doi: 10.1016/j.redox.2023.102972

Fig. 1.

Fig. 1

NSC34 cells expressing hSOD1G93A undergo early entrance into senescence. (A) Detection of EdU+ (red) in non-transfected (hSOD1-), hSOD1WT and hSOD1G93A transfected (hSOD1+, green) NSC34 cells. Scale bar, 20 μm. (B) Quantification of the proportion of EdU+ cells in NSC34 cells expressing hSOD1 within 3 weeks (n = 20–50 cells), *p < 0.05. (C-D) Representative and quantification of Western blot analysis of P53 protein level in NSC34 cells (n = 5, mean ± SEM, *p < 0.05, **p < 0.01). (E) Representative images of cellular senescence associated with β-galactosidase (SA-β-gal) staining in NSC34 cells. The black arrow indicated the SA-β-gal positive cells (green). Scale bar, 10 μm. (F) The proportion of SA-β-gal positive cells in total cells from each image field (n = 50–100 cells, mean ± SEM, **p < 0.01). (G) Immunofluorescence staining for hSOD1 (green), HMGB1 (red), and Hoechst (blue) in NSC34 cells. Scale bar, 10 μm. (H) The proportion of cells with cytoplasmic translocation of HMGB1 in the total hSOD1+ cells (n = 20 cells, mean ± SEM, *p < 0.05, **p < 0.01). (I–K) Representative and quantification of Western blot analysis of HMGB1 protein levels in the nucleus and cytosol of NSC34 cells (n = 5, mean ± SEM, *p < 0.05). (L) The proportion of thiol group level in total hSOD1 within NSC34 cells (n = 11, mean ± SEM, **p < 0.01).