Fig. 5: mESCs with differential STIP1 expression present changes in proliferation, DNA damage response and cell cycle profile and proteins.

A Representative immunofluorescence panel of cells stained with DAPI/Nuclei (red) and Ki67 (green). Scale bar = 10 µm. B Quantification of Ki67-positive (Ki67+) mESCs. Student’s t test with Welch’s correction, ****p < 0.0001, WT: n = 85; STIP1^ΔTPR1: n = 36. Mean ± SD. C Growth curve of WT and STIP1^∆TPR1 mESCs was analyzed per day by Student’s t test. Day 1 (24 h after seeding): **p = 0.0094; Day 3 (72 h): ***p = 0.0005; Day 5 (120 h): ***p = 0.0004; and Day 7 (168 h): ***p = 0.0004. Mean ± SD, n = 3. D Colorimetric analysis (corrected absorbance) of cell viability. Student’s t test, ***p = 0.009, Mean ± SD, n = 4. E Representative immunofluorescence panel of cells stained with DAPI/Nuclei (blue) and pH2AX (green). Scale bar = 10 µm. F Quantification of the number of pH2AX foci in each 10 µm. Student t test, ***p = 0.0009. Mean ± SD, n = 15. G Flow cytometry for WT and STIP1^∆TPR1 shown as representative scatter plots, staining for BrdU (S-phase) and 7-AAD (total DNA content). H Quantification of the percentage of cells for each cell cycle phase. Student’s t test, **p = 0.0085. Mean ± SD, n = 3. I Flow cytometry of cells labeled for RB1 (p < 0.0001, all comparisons), p53 (p < 0.0001, all comparisons), p21 (p < 0.0001, WT vs STIP1^TgA, STIP1^ΔTPR1 vs STIP1^TgA), CDC2 (p < 0.0001, all comparisons), cyclin B1 (p < 0.0001, all comparisons), and cyclin D1 (p < 0.0001, all comparisons). One-way ANOVA with Tukey’s post-hoc. Between 13.000–30.000 events were measured, and the mean and standard deviation of those events were used for statistics. The color pattern represents the respective groups across all charts.