Fig. 2: Ifih1 results in defects of the pluripotency and differentiation programme of ESCs.

a Two different ESC clones expressing a doxycycline-inducible N-terminal FLAG-tagged Ifih1 gene were used. Upon doxycycline stimulation, Ifih1 (MDA5) expression was confirmed by western blot (a) with anti-FLAG antibody. Tubulin serves as a loading control and expression was confirmed by RT-qPCR (b) of Ifih1 (t = 0 vs t = 8; MDA5(1) p = 0.000001; MDA5(2) p = 0.000002). 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. c Differentially upregulated and downregulated genes (abs log2FC > 0.4, p-val≤0.05) common to the two Ifih1-inducible clones were used for gene ontology analyses (biological process). d Shared upregulated genes for both clones were analysed using the Mouse Gene Atlas for predicted cell types. P-value is computed using the Fisher exact test as proportion test assuming a binomial distribution and independence of the genes. e Log2FC expression changes ± SD of neurogenesis-related genes following MDA5 induction in clones MDA5(1) and MDA5(2) were determined using RNA-seq data from three independent biological replicates. f Shared downregulated genes for both clones were analysed using the Mouse Gene Atlas for predicted cell types. Statistics as in (d). g Log2FC expression changes ± SD of pluripotency-associated genes following MDA5 induction in clones MDA5(1) and MDA5(2) were determined using RNA-seq data from three independent biological replicates.