Fig. 5: PDE5A overexpression in primary cultured mouse MSNs led to compromised dendrite arbor development and increased apoptosis.

A Western blot of PDE5A expression in control (NC) and PDE5A overexpressed (OE) MSNs. GAPDH was used as internal control. Quantification of PDE5A protein levels was calculated by relative gray value (Shapiro-Wilk test, n = 3 batches, ***p < 0.001). B Representative images of DIV14 MSNs expressing GFP and stained with MAP2 (a dendritic marker) from NC, OE, OE+tadalafil, and OE + DMSO MSNs, respectively. Scale bar, 20 μm. C Sholl analysis shows the number of dendritic intersections plotted against the distance from the center of the soma (2 way ANOVA Sidak’s multiple comparison test, p < 0.05 for 30–130 μm distance, p < 0.05 between NC and OE + tadalafil group in the range of 100–110 μm). D Sholl analysis shows total length of dendrites. 40 neurons each group from three independent batches were statistically analyzed (Shapiro-Wilk test,**p < 0.01 ***p < 0.001). E Representative images of DIV14 MSNs expressing GFP and stained with caspase 3 (an apoptosis marker) from the four groups. Scale bar, 150 μm. White arrow indicated apoptotic cells. F Ratio of caspase 3⁺ GFP⁺ cells in GFP⁺ cells was compared in the four groups (Shapiro-Wilk test, n = 3 batches, 3–4 fields per batch, ***p < 0.001).