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Fig. 6 | Cell & Bioscience

Fig. 6

From: Optogenetic stimulation inhibits the self-renewal of mouse embryonic stem cells

Fig. 6

Photoactivation triggers the differentiation of ChR2-GFP-V6.5 ESCs via ERK activation. a Hematoxylin and eosin staining of teratomas derived from ChR2-GFP-V6.5 ESCs in both the photoactivated group and the nonphotoactivated control group. Ectoderm, mesoderm and endoderm are marked with different color arrows. b qRT-PCR analysis of the mRNA level of lineage-specific genes in photoactivated ChR2-GFP-V6.5 ESCs and nonphotoactivated control cells. c Western blot analysis of the total ERK and p-ERK in ChR2-GFP-V6.5 ESCs after different intensities of optical stimulation (low intensity: 0.1 mW, medium intensity: 1 mW, high intensity: 10 mW). d Statistical analysis of the expression levels of ERK and p-ERK in photoactivated and nonphotoactivated ChR2-GFP-V6.5 ESCs. e Immunostaining for the pluripotency markers OCT4, SSEA-1, SOX2 and NANOG in photoactivated, photoactivated with MEK inhibitor PD0325901 (1 mM), nonphotoactivated and nonphotoactivated with MEK inhibitor PD0325901 (1 mM) ChR2-GFP-V6.5 ESCs. f qRT-PCR analysis of the pluripotency-associated genes Oct4, Sox2, Nanog, Esrrb and Klf4 in photoactivated, photoactivated with MEK inhibitor PD0325901 (1 mM), nonphotoactivated and nonphotoactivated with MEK inhibitor PD0325901 (1 mM) ChR2-GFP-V6.5 ESCs. g AP staining of in photoactivated, photoactivated with MEK inhibitor PD0325901 (1 mM), nonphotoactivated and nonphotoactivated with MEK inhibitor PD0325901 (1 mM) ChR2-GFP-V6.5 ESCs (n = 6, *P < 0.05, **P < 0.01, ***P < 0.001 compared to the nonphotoactivated control). h Summarize of calcium wave by photoactivation or KCl with or without extra-cellular calcium

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