Fig. 2: PHC1 is required for the pluripotency maintenance of hESCs.
From: PHC1 maintains pluripotency by organizing genome-wide chromatin interactions of the Nanog locus
a, b Morphology and colony-forming capacity of shScr-, shPHC1.1-, and shPHC1.2-infected hESCs. Scale bars, 600 μm. Cells were stained for alkaline phosphatase activity 14 or 18 days after plating. Bar plot shows mean colony-formation efficiencies normalized to the shScr. Mean ± s.d. of n = 3 independent experiments. Two-tailed unpaired t-tests were used (**p = 0.0022, *p = 0.0458). c Comparison of tumor sizes about 1 month after injection of shScr-, and shPHC1-infected hESCs into NOD/SCID mice. Data are presented as mean ± s.e.m. In each group, 4 NOD/SCID mice were injected. Two-tailed unpaired t-tests were used (*p = 0.0211). d WB analysis of PHC1, NANOG, OCT4, SOX2, RING1B, H2AK119ub1, and ACTIN protein levels in shScr and shPHC1-infected hESCs. e WB analysis of PHC1 and NANOG protein levels in hESCs infected with control and CRISPR-CAS9 vector targeting human PHC1 gene. f Immunofluorescent co-staining of PHC1 with NANOG, OCT4, or SOX2 in shScr- and shPHC1-infected hESCs. Arrowheads showed cells with low PHC1 and NANOG signals. Scale bars, 20 μm. g The ratios of cells exhibiting PHC1highNANOGlow and PHClowNANOGlow signals to the total number of stained cells in (f) were quantified. Mean ± s.d. of n = 3 independent counts for shPHC1.1 and shPHC1.2. Two-tailed unpaired t-tests were used (p = 0.0065 for shPHC1.1, p = 0.0048 for shPHC1.2). Source data are provided as a Source Data file.
