Fig. 2.

Snhg3 depletion impairs mouse embryonic stem cell (mESC) self-renewal. a mESCs were transfected with siControl or two different oligos targeting Snhg3 for 48 h and subjected to qRT-PCR to access knockdown efficiency. b Alkaline phosphatase (AP) staining of mESCs after siControl or siSnhg3 treatment; × 4 objective, scale bar = 100 μm. siControl- or siSnhg3-treated mESCs were re-plated onto feeders to form secondary ESC colonies for 7 days, which was followed by AP staining (c). The numbers of total ESC colonies were quantified in d. e siControl- or siSnhg3-treated mESCs were lysed for western blotting with indicated antibodies. GAPDH was used as a loading control. f Two stable Snhg3-knockdown mESCs were generated using a lentivirus. Knockdown efficiency was measured by qRT-PCR. g Cell cycle distribution of shControl or shSnhg3 cell lines was determined by flow cytometry. h Quantification of the percentage of cells in different cell cycle phases. i qRT-PCR analysis of cell cycle-related markers in shControl or shSnhg3 cell lines. Data are presented as mean ± SD; n = 3, two-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001 for all panels