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Fig. 2 | Epigenetics & Chromatin

Fig. 2

From: Maintenance of active chromatin states by HMGN2 is required for stem cell identity in a pluripotent stem cell model

Fig. 2

Expression of pluripotency markers in Hmgn knockout cells. a Relative expression of the pluripotency transcription factors Oct4, Sox2, and Nanog as determined by qRT-PCR. The graphs show the fold change relative to parental P19 cells (n = 3–10) (*p < 0.05, **p < 0.01, ***p < 0.001). b Western blot analysis of Nanog protein levels in whole cell extracts from parental P19, CON-a, N2-a, N2-b and N2-c cells. For both Actin and Nanog, all samples were run on the same gel and exposed at the same time. Actin is shown as a loading control. The relative ratios of Nanog to Actin, averaged from two replicate blots, are shown below. Immunofluorescence analysis of Nanog (c) and OCT4 (e) expression. Parental, N1-a and N2-a cells were fixed 24 h after seeding. DAPI was used to stain the nuclei (cyan). Scale bar indicates 50 µm. Nanog levels are heterogeneous, and cells with reduced expression are indicated by arrow heads. Images from all lines are shown in Additional file 1: Figure S4. FACS analysis of Nanog (d) and OCT4 (f) expression. E14-NSCs are the negative control. The bar graphs represent the relative fluorescence intensity (median) of each cell line, n = 3 (*p < 0.05, **p < 0.001, ***p < 0.0001). The cell count plots are representative examples to illustrate the distribution of the data for parental and knockout cells

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