Fig. 3

Role of Shp2 in primed pluripotency. A Light microscopic images of WT and KD ESCs at indicated times under LIF + 2i or bFGF/Activin culture condition (scale bars, 500 μm) (left), Graphical presentation of cell number at indicated time after seeding of 1 × 10⁴ ESCs (right). B Images of clonogenic assay of WT and KD ESCs at indicated time under LIF + 2i or bFGF/Activin culture condition (left), Graphical presentation of area ratio (A.U.) of clones at day 2 and day 4 determined by ImageJ (right). C Graphical illustration of competition assay with WT, labeled with red fluorescence [RF/GFP], and KD [GFP only] mESCs (top), Fluorescence microscopic images of WT or KD ESCs at indicated time after each culture condition, circles indicate KD ESCs [GFP only] (scale bars, 500 μm) (left), Relative fluorescence intensity of WT (red) and KD (green) ESCs from GFP images from 72 h was presented as a graph (right) (***p < 0.001, n = 26). D Immunoblotting analysis of WT and KD ESCs grown under bFGF/Activin for Shp2, pStat3, pErk1/2, Erk2 and β-actin. β-actin was used as a loading control