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. 2021 Apr 14;220(6):e202011014. doi: 10.1083/jcb.202011014

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© 2021 Vessoni et al.

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Figure S2. — Generation of isogenic iTERT hPSCs knocked out for TP53 (iTERT_TP53 KO) or both TP53 and RB1 (iTERT_TP53/RB1 KO). (A) DNA sequencing analysis confirming ablation of TP53 in the indicated cell lines. (B) DNA sequencing data analysis confirming ablation of Rb1 in the indicated cell lines. (C) Western blotting analysis of FLAG-TERT, p53, and pRb in iTERT hPSCs and their TP53 KO and TP53/Rb1 KO derivatives. GAPDH is shown as loading control. (D and E) Karyotype analysis of iTERT_TP53 KO and iTERT_TP53/Rb1 KO hPSCs, respectively. (F) Survival analysis of iTERT hPSCs, TP53 KO, and TP53/Rb1 KO after treatment with different concentrations of etoposide. Analysis performed by Sapphire700/DRAQ5 staining. (G) PDLs of iTERT hPSCs and fibroblasts, and their TP53 and TP53/RB1 KO counterparts, grown in the absence of DOX. (H) TRF analysis in iTERT, iTERT_TP53 KO, and iTERT_TP53/RB1 KO hPSCs grown in the absence of DOX for the indicated PDLs. Tel., telomere.