Fig. 4.

Genetic and pharmacologic activation of telomerase or inhibition of YAP1 suppresses immune activation pathways in IBD epithelium. (A) Micrograph of TIFs in CD duodenal organoids with or without overexpression of telomerase. (Scale bar, 10 µm.) (B) Quantification of TIFs in duodenal organoids from CD patient with or without overexpression of telomerase. Fifty cells from each group were quantified for TIFs. (C) Western blotting of lysates from the CD organoid line transduced with an inducible hTERT overexpression plasmid treated with or without (−DOX) doxycycline for 1 or 2 wk. (D) Western blotting of lysates from control (n = 2), CD (n = 2), and UC (n = 2) organoids with the indicated antibodies, indicating up-regulation of DNA damage and the ATM/YAP1/IL18 axis. (E and F) Western blotting for the indicated antibodies of lysates from older-onset CD organoids treated with or without TA-65 or NAD for 10 d (E) and quantification (F). (n = 2). (G and H) Western blotting for the indicated antibodies of lysates from older-onset UC organoids treated with or without TA-65 or NAD for 10 d (G) and quantification (H). (n = 2). (I and J) Western blotting with the indicated antibodies of lysates from YAP inhibitor treated or untreated CD organoids (I) and quantification (J). (n = 2). (K and L) Western blotting with the indicated antibodies of lysates from YAP inhibitor treated or untreated UC organoids (K) and quantification (L). (n = 2). n represents number of organoid lines used in the study. Each experiment was conducted at least two times.