Fig. 1: SGS iPSCs do not display of SETBP1 accumulation. | Nature Communications

Fig. 1: SGS iPSCs do not display of SETBP1 accumulation.

From: SETBP1 accumulation induces P53 inhibition and genotoxic stress in neural progenitors underlying neurodegeneration in Schinzel-Giedion syndrome

Fig. 1

a Fibroblast reprogramming from age-matched healthy donors (2) and SGS patients (2) and correction of patient-derived iPSCs (upper panel). Representative bright-field images (taken at the same magnification) of iPSC colonies derived from a healthy donor and SGS patients, (middle panel). Sanger sequencing confirmed the presence of the indicated mutations (lower panel, n = 6). b Back-mutation strategy in patients-derived iPSCs. For each patient, the mutation (green aa) was targeted with CRIPSR/Cas9, and the correction was confirmed by Sanger sequencing (correction efficiency: 1/48 clones for pt.1, 1/24clones for pt.2). Representative bright-field images (taken at the same magnification) of colonies from both isogenic corrected iPSC lines. c SETBP1 immunoblotting in isogenic control (D868D and I871I), mutant (D868N and I871T) and WT iPSCs and relative quantification. D868D vs. D868N P = 0.6973, I871I vs. I871T P > 0.9999; WT vs. MUT P = 0.9798. n = 3. d ET immunoblotting in isogenic control and mutant iPSCs and relative quantification. See Supplementary Dataset 3 for details of statistical analysis and Supplementary Fig. 1f for WT vs. MUT comparisons. n = 5. e Total and phosphorylated (pPP2A Tyr307) PP2A immunoblotting in isogenic control and mutant iPSCs and relative quantification of pPP2A/PP2A ratio. WT vs. D868D P = 0.8163; WT vs. D868N P = 0.9085; WT vs. I871I P = 0.4326; WT vs. I871T P = 0.9383; D868D vs. D868N P = 0.9994, I871I vs. I871T P = 0.8531; WT vs. MUT P = 0.8363. n = 4. f PP2A phosphatase activity in isogenic control and mutant iPSCs. D868D vs. D868N P = 0.9487, D868D vs. I871T P = 0.8743, WT vs. I871T P = 0.9882 n = 3. g Phosphorylated H3 (pH3 Ser10) immunostaining in isogenic control and mutant iPSCs and quantification. D868D vs. D868N P = 0.9998, I871I vs. I871T P = 0.9868, see Supplementary Fig. 1g for WT vs. MUT comparison. n = 4. All data expressed as mean ± SEM. ns: nonsignificant differences when P > 0.05. Images taken at the same magnification. c–g One-way ANOVA followed by Tukey post hoc test for multiple comparisons, except for WT vs. MUT comparisons two-sided unpaired t test in c and e.

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