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. 2024 Jun 10;10(12):e32680. doi: 10.1016/j.heliyon.2024.e32680

Fig. 4.

Fig. 4

Expression profile of REST and RCOR genes in human astrocytes. (A) Flow diagram for induced astrocyte generation. Representative images of iAs after 7 days (B), 14 days (C) and 21 days (D) of maturation. Scale bar = 100 μm. (E) Representative images of iAs stained for astrocyte marker, GFAP, at 7 and 14 days of maturation from iPSC2 cell line (Scale bar = 50 μm), (F) and the % positive cells for each target were calculated. Each data point represents the average of 3–4 fields of view per independent differentiation (n = 3 independent differentiations). (G) A glutamate uptake assay was completed on day 14 iAs derived from H9, iPSC1 and iPSC2 cell lines and determined by a serial dilution of glutamate standard (samples run in duplicate from 3 independent differentiations). (H) Day 14 iAs were incubated with Poly(I:C) and IL6 (encodes Interleukin-6) expression was analysed using one-step RT-qPCR to assess the ability to respond to inflammatory stimulation. (I) The expression levels of REST and RCOR genes was assessed in day 14 iAs derived from three hPSC cell lines using RT-qPCR. n = 3 independent differentiations, n = 3 technical replicates. Data was analysed using a Two-way ANOVA with a Holm-Sidak test for multiple comparisons, *p < 0.05, **p < 0.01, ***p < 0.001.