Fig. 7: SGS neurons inherit DNA damages and are prone to degenerate.

a Scheme for NPC-derived neuronal differentiation protocol (upper panel) and representative bright-field images (taken at the same magnification) of isogenic control (D868D) and mutant (D868N) neuronal cultures (lower panel). b Representative images (taken at the same magnification) of immunostaining for MAP2 (left) and relative quantification of isogenic control (D868D) and mutant (D868N) NPC-derived neuronal cultures. D868D vs. D868N *P = 0.0370; I871I vs. I871T P = 0.0768; D868N vs. I871T P = 0.5995. One-way ANOVA followed by Tukey post hoc test for multiple comparisons. n = 3. c Immunostaining and quantification for phosphorylated histone H2AX (pH2AX Ser139) in isogenic control (D868D) and mutant (D868N) NPC-derived neuronal cells. *P = 0.0107. Two-sided unpaired t test. n = 4. Images taken at the same magnification. d Representative images (taken at the same magnification) of immunostaining for MAP2 and PI and relative quantification of PI⁺ over MAP2⁺ cells in isogenic control and mutant NPC-derived neurons. ****P < 0.0001. Two-sided unpaired t test. n = 8. e Immunostaining and quantification for phosphorylated histone H2AX (pH2AX Ser139) in isogenic control (D868D) and mutant (D868N) iPSC-derived neuronal cells. P = 0.7200. Two-sided unpaired t test. n = 4. Images taken at the same magnification. f Representative images of immunostaining for MAP2 and PI and relative quantification of PI⁺ over MAP2⁺ cells in isogenic control and mutant iPSC-derived neurons. P = 0.2337. Two-sided unpaired t test. n = 3. Images taken at the same magnification. All data expressed as mean ± SEM. ns: nonsignificant differences when P > 0.05; *P < 0.05, ^∗∗∗∗P < 0.0001. Analysis was performed at 4 weeks of neuronal differentiation.