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. 2022 May 26;119(22):e2123202119. doi: 10.1073/pnas.2123202119

Fig. 5.

Fig. 5.

Notch1 is a direct target of p73α1 and highly expressed in inflamed E12+/− mouse tissues. (A) The level of NOTCH1, HES1, and HPRT1 transcripts was measured in isogenic control, TAp73−/−, ΔNp73−/−, and E12−/− H1299 cells. (Right) qPCR was used to analyze relative NOTCH1 and HES1 expression. One-way ANOVA was used to calculate P values. **P < 0.01; ****P < 0.0001. (B) The level of TAp73α, Notch1 NTM, NICD, Hes1, and vinculin proteins was measured in isogenic control, TAp73−/−, ΔNp73−/−, and E12−/− H1299 cells. (C) The level of p73α, p73α1, NOTCH1, HES1, and HPRT1 transcripts was measured in isogenic control and E12−/− H1299 cells transiently transfected with Scr or si-E11#1 for 3 d. (Right) qPCR was used to analyze relative NOTCH1 and HES1 expression. One-way ANOVA was used to calculate P values. ***P < 0.001; ****P < 0.0001. (D) ChIP analysis was performed with isogenic control and E12−/− H1299 cells. Cell lysates were immunoprecipitated with control IgG or anti-TAp73 overnight to bring down the DNA-protein immunocomplex, and the DNA fragments were visualized by PCR with primers for NOTCH1, CDKN1A, and GAPDH promoters. (E) The levels of Notch1, Hes1, and Actb transcripts were measured in liver tissue from age- and sex-matched WT and E12+/− mice (100 wk; Female). (Right) qPCR was used to analyze relative Notch1 and Hes1 expression. One-way ANOVA was used to calculate P values. **P < 0.01; ***P < 0.001; ****P < 0.0001. (F) The level of Trp73α, Notch1, Hes1, and Actb transcripts was measured in salivary gland tissue from age- and sex-matched WT and E12+/− mice (100 wk; Female). (Right) qPCR was used to analyze relative Notch1 and Hes1 expression. **P < 0.01; ***P < 0.001; ****P < 0.0001. One-way ANOVA was used to calculate P values. (G) The level of Trp73α, Trp73α1, Notch1, Hes1, and Actb transcripts was measured in spleen tissue from age- and sex-matched WT and E12+/− mice (100 wk; Male). (Right) qPCR was used to analyze relative Notch1 and Hes1 expression. One-way ANOVA was used to calculate P values. *P < 0.05; ***P < 0.001.