Figure 2. lincRNAs are critical for the maintenance of pluripotency.
(a) Activity from a Nanog promoter driving luciferase, following treatment with control hairpins (black) or hairpins targeting luciferase (green), selected protein-coding regulators (red), and lincRNAs (blue). (b) Relative mRNA expression levels of Oct4 following knockdown of selected protein-coding (red) and lincRNA (blue) genes affecting Nanog–luciferase levels. The best hairpin (black line) and second best hairpin (grey line) are shown. All knockdowns are significant with a p-value<0.01. Error bars represent standard error (n=4). (c) Morphology of ESCs and immunofluorescence staining of Oct4 for a negative control hairpin (black line), and hairpins targeting Oct4 (red line), and two lincRNAs (blue line). The first row shows bright field images, the second row shows immunofluorescence staining of the Oct4 protein, and the third row shows DAPI staining of the nuclei.