Fig. 5

Silencing of Dppa3 downregulated Nanog expression. A Morphology of control ES cells and Dppa3 knockdown ES cells. Scale bar = 100 μm. B Left panel: Western blot analysis of Oct4, Nanog and Dppa3 protein levels in control and Dppa3 knockdown ES cells. Right panel: Quantification of Oct4, Nanog and Dppa3 protein levels normalized to Tubulin by the ImageJ software. The data are presented as mean ± SEM. (n = 3; *p < 0.05, **p < 0.01). C. Left panel: Immunofluorescence images of Nanog expression in control and Dppa3 knockdown ES cells. Scale bar = 100 μm. Right panel: Intensity analysis of every specific cell by image J. D Flow cytometry analysis of Nanog positive cells in control ES cells and Dppa3 knockdown ES cells. E Left panel: The protein levels of Nanog in control and Dppa3 knockdown ES cells were analyzed at the indicated time points (0 h, 1 h, 2 h, 3 h) after MG132 treatment (20 mM). Right panel: Quantification Nanog protein levels normalized to Tubulin by ImageJ software. The data are presented as mean ± SEM. F Left panel: Nanog protein levels were analyzed at the indicated time points (0 h, 1 h, 2 h, 3 h) after CHX treatment (2 mM). Right panel: Quantification of Nanog protein levels normalized to Tubulin by the ImageJ software. The data are presented as mean ± SEM. (n = 3; *p < 0.05, **p < 0.01)