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. 2021 Mar 1;131(5):e143173. doi: 10.1172/JCI143173

Figure 6. Endogenous YAP interacts with TFEB in CMs transduced with Ad-sh-RagA/B and promotes TFEB-mediated transcription.

Figure 6

(A and B) CMs were transduced with or without Ad-sh-RagA/B. (A) Representative immunoblots of cytosolic and nuclear fractions. Blots run in parallel, contemporaneously, using identical samples are shown. n = 5. (B) CM lysates were subjected to immunoprecipitation with anti-YAP or control antibody. Original lysates (input) and the immunoprecipitation samples were then subjected to immunoblot analyses. Representative images are shown. For input, a blot run in parallel, contemporaneously, using identical samples is shown. n = 5. (C) Triple immunostaining with anti-YAP and anti-TFEB antibodies and DAPI of CMs transduced with Ad-sh-RagA/B. n = 6. (D) PLAs using anti-YAP and anti-TFEB antibodies in CMs transduced with Ad-sh-RagA/B. Inset shows a 3.2-fold magnification of area indicated by yellow rectangle. n = 3. (E) TFEB reporter gene assays were conducted with neonatal CMs transduced with Ad-sh-Scr or Ad-sh-YAP in the presence or absence of Ad-sh-RagA/B. n = 6. (F) ChIP assays with anti-YAP antibody in CMs transduced with Ad-sh-Scr or Ad-sh-YAP in the presence or absence of Ad-sh-RagA/B. The precipitated chromatin was subjected to PCR to detect the presence of the TFEB-binding element in the rat MAPLC3B promoter, as indicated in the inset. n = 6. Results are expressed as mean ± SEM. *P < 0.05; **P < 0.01, ANOVA.