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. 2020 Mar 16;10:4829. doi: 10.1038/s41598-020-61640-9

Figure 5.

Figure 5

ATRA supresses KRT1 and KRT10 expression while KRT2 expression is increased. NHEK (200,000 cells) were mixed with 106 irradiated 3T3 fibroblasts in 2 ml of CS-FCS containing RM+ with or without PR per well of a 12 well plate in triplicates for each ATRA concentration for 3 days. ATRA stock was made in DMSO and further diluted in ethanol (EtOH). The cells were grown in three different ATRA concentrations of 1 µM, 2 µM and 3 µM and DMSO/EtOH was used as a vehicle control (0.003%/0.03% maximum) for 24 h after which the feeder was removed by squirting with PBS/EDTA and lysates were collected for qPCR analysis for (A) KRT1, (B) KRT10 and (C) KRT2. Data are shown as fold expression normalised to the expression of two housekeeping genes, POLR2A and YAP1. Statistical analyses: n = 3, Error bars =SEM. One-way ANOVA (shown by a horizontal line over each graph) to measure the p values at different concentrations of ATRA compared to the control (no ATRA). Two-way ANOVA was used to measure the statistical significance between PR+ and PR- groups for each keratin mRNA (KRT1, KRT10, and KRT2), p values are given by asterisks (ns = p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001).