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. 2014 Jul 7;35(8):1023–1030. doi: 10.1038/aps.2014.51

Figure 2.

Figure 2

Activation of SSTR2 prevents mESC differentiation caused by LIF deprivation. (A) Morphology and alkaline phosphatase (AP) staining of E14 cells cultured in mES media containing LIF (1000 U/mL) or mES media without LIF but supplemented with 2i (1 μmol/L PD0325901 and 3 μmol/L CHIR99021) or the SSTR2 agonist octreotide (1 μmol/L) (Scale bar: 50 μm). (B) The percent of undifferentiated colonies (according to AP staining and morphology) of E14 cells cultured in mES media containing LIF, 2i, octreotide or seglitide (SSTR2 agonist) at various concentrations for 3 d. The data are the mean±SEM (n=3). cP<0.01 vs LIF(−) group. (C) Quantitative RT-PCR analysis of pluripotency genes and SSTR2 in cells corresponding to (A). Data are the mean±SEM (n=3). bP<0.05, cP<0.01 vs LIF(+) condition. eP<0.05, fP<0.01 vs LIF(−) condition. (D) Immunofluorescence staining of pluripotency markers (Oct4, Nanog, and SSEA1) in mESCs cultured in octreotide (1 μmol/L)-containing, LIF-free media. (Scale bar: 10 μm). (E) AP staining of E14 cells pre-incubated with various concentrations of the SSTR2 antagonist S4 for 24 h and then cultured in LIF-free media containing octreotide (1 μmol/L) (Scale bar: 50 μm). (F) The percent of undifferentiated colonies (according to AP staining and morphology) of E14 cells cultured in the indicated media. cP<0.01 vs LIF(−) condition. fP<0.01 vs octreotide alone.