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. 2020 Nov 10;9(11):2451. doi: 10.3390/cells9112451

Figure 1.

Figure 1

Neuronal conversion of hESC-derived GPCs in 26 days: (A) Schematic of the reprogramming strategy using the ADLSF factor combination with or without REST inhibition. (B) RT-qPCR analysis showed downregulation of glial markers and upregulation of neuronal genes at day 26 after transduction. Kruskal–Wallis test, Dunn’s multiple comparisons test (n = 6–8 for CTRL GM, n = 7–8 for CTRL NDiff, n = 6–8 for ADLSF, and n = 3–4 for ADLSF + shREST): *** p < 0.005; * p < 0.05. (C,D) PDGFRα/GFAP and TAU immunostainings of (C) reprogrammed neurons generated with ADLSF and ADLSF + shREST and (D) control glial cells kept in glial medium (CTRL GM) or neuronal conversion medium (CTRL NDiff). (E) Quantification of immunodetected TAU+ cells. Two-tailed unpaired t-test with Welch´s correction (n = 11 for ADLSF and n = 4 for ADLSF + shREST): * p = 0.002; df = 11.97. (F) Quantification of neurite profile in TAU+ induced neurons. No significative difference (ns) was detected when comparing ADLSF (n = 11) and ADLSF + shREST (n = 5) using the two-tailed unpaired t-test. Data are presented as mean ± SEM. Each data point represents a replicate from an independent experiment. For the measurement of neurite length, each datapoint was normalized to the mean of the ADLSF condition. Scale bars: (C,D) 100 μm. Abbreviations: SM, small molecules; GF, growth factors; DOX, doxycycline.