Extended Data Fig. 1: Healthy control and HD patient fibroblasts can be directly reprogrammed into MSNs.
a. Reprogrammed cells by transduction of miR-9/9*-124+CDM immunostained for neuronal markers, TUBB3, and MSN marker, DARPP32 at PID30. Scale bars 20 μm. b. Expression of each CDM factor in reprogramming cells by immunostaining for CTIP2, DLX1, DLX2, MYT1L, and TUBB3. Non-transduced fibroblasts were used as a negative control for immunostaining. Scale bars 20 μm. c. The neuronal morphology across all samples we used in the current study stained positive for TUBB3 successfully undergo direct conversion by miR-9/9*-124+CDM. Scale bars 20 μm. d. Left, representative images of healthy control Young, Old, Pre-HD, and HD MSNs marked by TUBB3. Images processed by CellProfiler to identify neurites and associated cell soma. Scale bars, 100 μm. Right, the measurement of mean neurite length and mean number of neurite branches in reprogrammed Healthy control Young, Old, Pre-HD, and HD-MSNs at PID21 and PID35 (n = 24 individual’s reprogrammed MSNs). Statistical significance was determined using one-way ANOVA, ****p < 0.0001, ***p < 0.001 (Old vs. HD in neurite length p = 0.0007, Old vs. HD in neurite branches p = 0.0001, Pre-HD vs. HD in neurite branches p = 0.0002), **p < 0.01 (Young vs. HD in neurite length p = 0.0012, Pre-HD vs. HD in neurite length p = 0.0026), ns, not significant. Mean±s.e.m.
