fig3

Figure 3. GSNOR loss of function alters S-nitrosylation and expression of GSK3β. (A) qRT-PCR analysis of Gsk3β in murine iPSC^WT (white circles) and iPSC^GSNOR-/- (black circles) during cardiomyocyte differentiation. (B) S-nitrosylation of Gsk3β in murine iPSC^GSNOR-/- and iPSC^WT as assessed by SNO-RAC on Day 4. Effects on Gsk3β mRNA levels, in (C) murine iPSC^WT, following 3 h incubation with 3 different concentrations of the NO donor, GSNO, on Day 4, and in (D) hiPSCs (brown circles), following 24 h of incubation with GSNO beginning on Day 1. (E) Western blot analysis of Axin 1 and Hsp90 (internal control) in murine iPSC^WT (white circles) and iPSC^GSNOR-/- (black circles) on Day 0, 2, 4 and 6 of cardiomyocyte differentiation. qRT-PCR analysis of Axin 1 (F) and Axin 2 (G) in murine iPSC^WT (white circles) and iPSC^GSNOR-/- (black circles) during cardiomyocyte differentiation. Data are the mean ± SEM of 3 independent experiments. P-values were calculated using Student’s t-test at each time point. *P < 0.05.