Skip to main content
. 2022 Oct 17;132(20):e160152. doi: 10.1172/JCI160152

Figure 4. ChAT expression is activated transcriptionally through YAP mediation.

Figure 4

(A) Microscopic image showing the uncloned PC9 cells or cells from 5 different single-cell-derived PC9 clones treated with 2 μM osimertinib for 9 days to identify DTPs. (B) Quantification of ACh levels in 5 different single-cell-derived PC9 clones treated with 2 μM osimertinib for 9 days (n = 5). (C) Protein levels of the short ChAT isoform in 5 different single-cell-derived PC9 clones treated with 2 μM osimertinib for 9 days. (D) ChIP-qPCR analyses in PC9 cells showing YAP bound to the enhancers 1 and 3 of the ChAT gene after 2 μM osimertinib treatment for 48 hours (n = 4). (E) Luciferase reporter assay in PC9 cells with stable silencing of YAP or control shRNA (shCtrl) treated with 2 μM osimertinib for 72 hours (n = 3). (F) Relative ChAT mRNA levels in PC9 cells with shYAP or shCtrl treated with 2 μM osimertinib for 9 days. Gene silencing was confirmed by Western blot on the left. (G) Relative protein levels of the short ChAT isoform in PC9 cells with shYAP or shCtrl treated with 2 μM osimertinib for 9 days. (H) Quantification of ACh levels in PC9 cells with shYAP or shCtrl treated with 2 μM osimertinib for 9 days (n = 5). In B, D–F, and H, data are shown as mean ± SEM. Significance was assessed using 2-way ANOVA with Tukey’s test (B, D, E, and H) or 1-way ANOVA with Tukey’s test (F).