Fig. 3.

Trincr1 promotes ESC self-renewal and suppresses ERK target genes. a RT-qPCR analysis of pluripotency genes and Fgf5 in Trincr1−/− ESCs in PD + LIF condition. The β-actin gene was used as a control. For each gene, data were normalized to the mRNA level of wild-type ESCs. n = 3 biological replicates. b Colony formation assay for wild type, Trincr1−/− ESCs cultured in 2i + LIF or PD + LIF condition on feeder cells in a 12-well plate. Two independent Trincr1−/− ESC clones were analyzed. Data were normalized to the colony number of wild-type ESCs in 2i + LIF. n = 3 biological replicates. c Principal component analysis of all protein-coding genes in wild type and Trincr1−/− ESCs in 2i + LIF and PD + LIF. d KEGG pathway analysis of upregulated genes in Trincr1−/− ESCs in PD + LIF. Top 8 enriched pathways are shown with P values and fold of enrichment. e GSEA analysis for MAPK/ERK targets in wild type and Trincr1−/− ESCs in PD + LIF. The distinct peak at the end of the ranked list indicates that MAPK/ERK target genes are generally upregulated in Trincr1−/− ESCs. Shown are mean ± SD. For a, P values were determined by unpaired two-sided Student’s t-test. For b, P values were determined by unpaired two-way ANOVA with two-sided Dunnett’s test. For e, P value was determined by an empirical gene set-based permutation test