Supplementary Figure 3: ChIP-qPCR analysis of REST targets and characterization of calcium influx in response to membrane depolarization in induced neurons with shPTB plus various neuronal lineage-specific transcription factors.
From: Sequential regulatory loops as key gatekeepers for neuronal reprogramming in human cells
(a) ChIP-qPCR analysis of REST binding at the promoters of indicated neuronal transcription factors and miRNA loci in HAFs. Two-tailed unpaired Student’s t-test was applied to calculate significance. Data are presented as mean± SD. n=3 independent experiments. Ascl1: υ=4, t=5.336, P=0.00594; Myt1l: υ=4, t=11.605, P=0.000315); NeuroD1: υ=4, t=34.602, P=0.000004; Olig2: υ=4, t=12.332, P=0.000248; Brn2: υ=4, t=3.737, P=0.020; miR-124: υ=4, t= 5.812, P=0.00436. *P<0.05, **P<0.01, ***P<0.001.
(b) Screening for transcription factors capable of promoting MAP2 expression (Green) when combined with PTB knockdown in HAFs.
(c) Calcium influx in neuronal cells induced by co-expression of shPTB and BRN2 in HAFs. Scale bar: 30 µm.
