Fig. 1: Ifih1 expression is induced upon differentiation.
From: Double stranded RNA sensing is silenced during early embryonic development
Expression of genes involved in TLR signalling, RLR signalling pathway and JAK/STAT signalling in ESCs differentiating to ectoderm (a), mesoderm (b) and endoderm (c), as obtained from RNA-seq. Samples were taken at successive days after starting differentiation (day 0 corresponds to ESCs, until day 6 or 9 of differentiation). Expression is calculated as normalised counts and relative to ESCs (day 0). RT-qPCR analyses for selected pluripotency genes (d), differentiation markers (Nanog p = 0.000001; Pou5f1 p = 0.001; Sox17 p = 0.001; Foxa2 p = 0.0007) (e) and RLR signalling pathway genes (Ifih1 p = 0.0001; Rigi p = 0.006; Irf3 p = 0.01; Irf7 p = 0.05; Mavs 0.04) (f) after embryoid body differentiation (Diff) of pluripotent ESCs (ESC). Data in d–f represent the average of three biological replicates ± the standard error of the mean (SEM). Single factor ANOVA was used to calculate significant differences amongst comparisons, followed by an F-test for variance and appropriate two-tailed t-test. g RT-qPCR analyses of Ifnb1 (ESC p = 0.006; diff p = 0.03), Tnf (ESC p = 0.002; diff p = 0.0004) and Cxcl10 (ESC p = 0.02; diff p = 0.001) upon dsRNA stimulation with poly(I:C) (p(I:C)) of ESCs and in vitro differentiated ESCs (as in d). Replicates and statistical tests as in (d). h Both ESCs and in vitro differentiated cells (as in d) were stimulated with exogenous IFN-β (100 pg/ml), ISGs induction was quantified by RT-qPCR. (Stat1 diff p = 0.006; Oas1 diff p = 0.000004; Isg15 ESC p = 0.03 diff p = 0.0001). Replicates and statistical tests as in (d). i ESCs were stimulated with exogenous IFN-β (100 pg/ml), and ISG induction, including Ifih1(t = 0 vs t = 6; p = 0.0004), Oas1 (t = 0 vs t = 8; p = 0.01) and Stat1 was measured by RT-qPCR. Data represent the average of three biological replicates ± SD. One-way ANOVA was used to calculate significant differences amongst comparisons, followed by an F-test for variance and Tukey HSD. *p-val ≤ 0.05, **p-val ≤ 0.01, ***p-val ≤ 0.001.
