Figure 3.

Linc1614 mainly mediates Sox2 function in the repression of developmental genes. (A) Representative images following the overexpression of Sox2 in sh1614 ESCs. The cell morphology under bright-field microscopy (top) and AP-stained colonies (bottom) are shown. Scale bar, 100 μm. (B) qPCR analysis showed the restoration of Sox2 expression in linc1614-knockdown ESCs. Data were presented as mean ± SD (n = 3). Student’s t-test was performed for significance (*^,#P < 0.05 and **P < 0.01). * relative to Ctrl and ^# relative to sh1614 + luc. (C) Statistical analysis of the colony morphology for the overexpression of Sox2 in sh1614 ESCs. More than 250 individual colonies were counted in each of the three independent experiments. (D and E) The expression of pluripotency genes upon overexpression of Sox2 in sh1614 ESCs was analyzed by qPCR (D) and western blotting (E). (F) qPCR analysis of the expression of developmental genes upon overexpression of Sox2 in sh1614 ESCs. Data were presented as mean ± SD (n = 3). Student’s t-test (*P < 0.05, **P < 0.01, and ***P < 0.001) was performed relative to Ctrl. n.s. represented no significance. (G) Pairwise comparisons of the number of genes showing changes in regulation (>1.5-fold change compared with that of the control cells) in sh1614 and shSox2 ESCs. The numbers of upregulated (red) and downregulated (green) genes are shown. (H) GO term analysis for genes that were differentially expressed in shSox2 ESCs compared with control ESCs. GO enrichment was assessed using the DAVID gene ontology functional annotation tool. The 10 most significant GO terms are shown. (I) GO term analysis of genes that were differentially expressed in sh1614 ESCs compared with control ESCs. (J) Heat maps of the overlap of the differentially expressed genes following knockdown of Sox2 or linc1614 are shown. The scale indicated the fold change compared with control ESCs. (K) GO term analysis of upregulated and downregulated genes in both shSox2 and sh1614 ESCs are shown.