Fig. 6

CHIR99021 + PD0325901 enhanced pluripotency in GSPCs and ES-like cells. a-d The morphology of GSPCs cultured in medium 2 (a), or in medium 7 for 1 passage (b), 4 passages (c) and 10 passages (arrowheads indicate some recovered colonies with compact structure) (d) was exhibited. e The morphology of ES-like cells cultured in medium 8 for 10 passages was exhibited. f-k IF staining assays were used to detect the expression of NANOG in GSPCs cultured in medium 7 (f, NANOG. g, DAPI. h, Merge) and ES-like cells cultured in medium 8 for 10 passages (i, NANOG. j, DAPI. k, Merge). m-p The morphologies of ES-like cells cultured in medium 3 (m), or medium 9 for 1 passage (n), 6 passages (arrowheads indicate some recovered colonies with compact structure) (o) and 10 passages (p), were exhibited. q-s Expression of NANOG in ES-like cells cultured in medium 9 was detected (q, NANOG. r, DAPI. s, Merge). t Expression levels of NANOG, OCT4, MVH and β-tubulin in ES-like cells cultured in medium 3 or medium 9 for 10 passages were determined using Western blot. u-v Expression levels of NANOG, SOX2, MVH, PLZF and β-tubulin in GSPCs (u) and ES-like cells (v) cultured with CHIR99021, PD0325901 or CHIR99021 + PD0325901 were determined using Western blot. w-x Growth curves of GSPCs (w) and ES-like cells (x) cultured with CHIR99021, PD0325901 or CHIR99021 + PD0325901 were exhibited. Scale bar = 20 μm, data represent as mean ± SD *p < 0.05; **p < 0.01