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. 2021 Jul 2;12:4090. doi: 10.1038/s41467-021-24373-5

Fig. 2. SS18 Regulates cJUN-induced PST.

Fig. 2

a Construct design for SS18 knockout in mESC. The cleavage sites of Cas9 were designed flanking exon1 and exon2 within Ss18 loci. b SS18KO mESCs are similar to WT ones in morphology. Three biological replicates. c Gene expression profiles by RNA-seq for WT and SS18 KO mESCs. SS18 knockout has little impact on most pluripotent genes but delays the c-JUN induced PST process significantly. d Rescue of the delayed PST as indicated by Nanog expression in SS18 KO mESCs with SS18 in lentiviral vector. Data are mean ± s.d., two-sided, unpaired t test; n = 3 independent experiments, **p < 0.01. e The experimental design to test the window of sensitivity for SS18 response to c-Jun-induced PST. f Results from (e) to show that OCT4-GFP positive colony numbers recovered for 48 h in 2i/LIF medium after c-Jun induction at the indicated time points for wild type and SS18 KO ESCs. Data are mean ± s.d., two-sided, unpaired t test; n = 3 independent experiments. **p < 0.01, ****p < 0.0001. Note no change for 4 h and the greatest change at 6 h for the window of sensitivity. g Representative images for the c-Jun-induced PST in WT, Ss18−/− and Ss18−/− overexpressing Ss18, respectively. Three biological replicates.