Fig. 1 | Cell Death & Differentiation

Fig. 1

From: Caspase-8, receptor-interacting protein kinase 1 (RIPK1), and RIPK3 regulate retinoic acid-induced cell differentiation and necroptosis

Fig. 1

Knockdown of Casp8 expression in ES cells enhanced RA-induced cell differentiation. a Dox-inducible (Tet-On) shCasp8-expression system. TetR, tetracycline repressor; and U6tet, mouse U6 promoter joining the tetracycline operator. b Validation of induced knockdown of Casp8 expression in Tet-On shCasp8 ES cells by western blot analysis after treatment with 1 μg/ml Dox for indicated days. Molecular weight markers are indicated (kDa). Neuronal differentiation of Dox (1 μg/ml)-treated or -untreated Tet-On shCasp8 and Tet-On shGFP ES cells was analyzed by phase-contrast microscopy (c) or fluorescence microscopy (d) after 6-day formation of EBs. EBs were treated with or without 1 μM RA for last 4 days. Scale bars, 200 μm. Cells were stained with anti-Tuj1 antibody in d. e qRT-PCR analysis of Oct3/4 was carried out using EBs defined in c. f qRT-PCR analysis of Nestin, Tuj1, and Casp8 was carried out using ES cells at the indicated times after formation of EBs defined in c. **p < 0.01, *p < 0.05 and n.s.d. (no significant difference: p > 0.95)

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