Extended Data Fig. 5: Effect of NF-κB knockdown, H2O2, TNFα and the overexpression of Ascl1 on the reprogramming of fibroblasts to CiPCs.
From: Pharmacologic fibroblast reprogramming into photoreceptors restores vision
a, qPCR analysis following shRNA-mediated knockdown (kd) of RelA (which encodes NF-κB) in MEFs. b, qPCR analysis of Ascl1 expression after the reprogramming of RelA-depleted MEFs on day 11. c, Number of Nrl–GFP cells after the conversion of RelA-knockdown MEFs on day 11. d, Effect of different concentrations of H2O2, generated by galactose and galactose oxidase (GAO), on the reprogramming of fibroblasts to CiPCs. e, Effect of cytokine (TNF, 50 ng ml−1) on CiPC conversion. f, qPCR analysis showing the overexpression (OE) of Ascl1 in MEFs. g, Phase contrast micrographs of MEFs overexpressing Ascl1 before and after reprogramming in inducing medium. No Nrl–GFP+ cells were evident up to day 15. h, Fluorometric measurement of mROS in Ascl1-overexpressing cells on day 8 of reprogramming. Data are presented as mean ± s.e.m. of n = 3 independently treated wells. Statistical significance was assessed using a two-tailed Student’s t-test.
