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. 2019 Apr 4;13:120. doi: 10.3389/fncel.2019.00120

Figure 5.

Figure 5

Alterations in Drp1 levels and function affect neuronal differentiation induction. Drp1wt, Drp1K38A, Drp1 shRNA and control P19 stable clones were induced to differentiate with RA. RNA was extracted from d1 to d6 and used to analyze Mash1, Wnt1 and N-cadherin (d1–d4; A), Oct3/4 (d1–d4) and Sox2 (d1–d6) (B), and FGF8 (d1–d4) expression levels (C) by Real-Time PCR. Undifferentiated P19 cells stably transfected with the vector alone were used as an endogenous control (set at 1). Results are expressed as fold increase of endogenous control and reported as individual data plus the mean ± SEM (two-way ANOVA followed by Tukey’s multiple comparison test, n = 8). * vs ctr (***p < 0.001, **p < 0.01); + vs Drp1wt (+++p < 0.001, ++p < 0.01). Lines indicate samples to whom asterisks are referred. (D) Total extracts were prepared from Drp1wt, Drp1K38A, Drp1 shRNA and control clones 24 h after RA addiction, run on a 10% SDS-PAGE gel and probed with anti phospho-Akt (p-Akt), Akt, phospho-Ser9-GSK3β (p-GSK3β), GSK3β, phospho-ERK1/2 (p-ERK), ERK1/2, phospho-Ser206-Smad1 (p-Smad1) and Smad1 Abs. Uncropped gels are in Supplementary Figure S10. Quantification is reported in Supplementary Figure S11.