a, RCAN1 expression in fibroblasts transduced with shRCAN1 (top) or RCAN1 (middle) in a dose-dependent manner. RCAN1 expression in HD-MSNs (HD.43) transduced with shCtrl, shRCAN1, or RCAN1 (bottom). b, Representative image (top) and quantification (bottom) of DARPP-32-positive cells from four independent HD-MSNs transduced with shCtrl, shRCAN1, or shCaN (HD.43, HD.40, HD.47, HD.45). Cells were immunostained with anti-DARPP-32 and TUBB3 antibodies. An average of 183 cells of each were counted from three or more randomly chosen fields (n = 4). Scale bars represent 10 μM. c, RT-qPCR analysis of the expression of RCAN1 and CaN in (b) (n = 12, 12, 8, 8). Statistical significance was determined using one-way ANOVA with Tukey’s post-hoc test (b) and two-tailed unpaired t-test (c); ****p < 0.0001, ns, not significant, and mean±s.e.m (b,c). The sample size (n) corresponds to the number of biologically independent samples (b,c).