Fig 2: Ablation of transforming growth factor β receptor II gene Tgfbr2 (r2) in myofibroblasts does not alter misfolded protein accumulation in mutant αB‐crystallin (CryABR 120G) hearts. A, Immunofluorescent staining of CryAB (green) with α‐actinin (red) shows no difference in aggregate accumulations in cardiomyocytes derived from Cry ABR 120G /Postnmcm/r2 f/f hearts with (+) or without (−) r2 ablation in the myofibroblasts (n=4). B and C, Western blot showing CryAB protein levels present in soluble and insoluble (aggregate‐containing) fractions. D and E, Quantitation of CryAB protein expression in soluble and insoluble fractions derived from hearts of the different experimental cohorts as indicated. CryAB was not detected in fractions derived from animals not expressing the CryABR 120G. Comparisons were made using 1‐way ANOVA followed by Tukey's post hoc test (n=4). *P<0.05 vs Cre induced Cry ABR 120G /Postnmcm/r2 f/f. GAPDH indicates glyceraldehyde 3‐phosphate; Ntg, nontransgenic.

Fig 3: Axl reduction in PCa cells suppresses the cellular dormancy of PCa cells in bone marrow.(A) Tumor growth monitored by bioluminescence imaging (BLI) in marrow. PCa sh Control and sh Axl cells labeled with luciferase were injected into tibiae of SCID mice (n = 5). (B) Immunohistochemistry showing Ki-67 expression in PCa sh Control and PCa sh Axl cells in BM. PCa cells were recognized by pan cytokeratin with green color, and Ki-67 was shown with red color. Scale bar, 20 μm. Ki-67 negative and positive PCa cells were counted and the percentage of Ki-67 negative PCa cells was shown. *p < 0.05 compared to PC3 sh Control cells. #p < 0.05 compared to DU145 sh Control cells. (C) Immunohistochemistry showing TGFGBR2 expression in PCa sh Control and PCa sh Axl cells in BM. PCa cells were recognized by pan cytokeratin with green color, and TGFBR2 was shown with red color. Scale bar, 10 μm. TGFBR2 positive and negative PCa cells were counted and the percentage of TGFBR2 positive PCa cells was shown. *p < 0.05 compared to PC3 sh Control cells. #p < 0.05 compared to DU145 sh Control cells. (D) Immunohistochemistry showing TGFGBR3 expression in PCa sh Control and PCa sh Axl cells in BM. PCa cells were recognized by pan cytokeratin with green color, and TGFBR3 was shown with red color. Scale bar, 10 μm. TGFBR3 positive and negative PCa cells were counted and the percentage of TGFBR3 positive PCa cells was shown. **p < 0.01 compared to PC3 sh Control cells. N.D. means not detected.

Fig 4: Exosomal miR-671 alleviates fibrosis and cell apoptosis in myocardial tissues in mice with MI. a miR-671 expression in murine myocardial tissues nearby the infarcted area detected using RT-qPCR; b Protein levels of TGFBR2 and p-Smad2 in murine myocardial tissues nearby the infarcted area examined by IHC staining; c Concentrations of IL-6 and TNF-α in the homogenate of murine myocardial tissues examined by ELISA kits; d Myocardial fibrosis in mouse examined by masson's trichrome staining; e Cell apoptosis in murine myocardial tissues examined by TUNEL assay. N = 8 in each group; representative images are provided. Data were collected from three experiments and exhibited as mean ± SEM. Differences were analyzed by one-way ANOVA (a, d, and e) or two-way ANOVA (b, c); *p < 0.05 vs. sham group; #p < 0.05 vs. Exo-NC group.

Fig 5: MYEOV attenuates miRNA-mediated suppression of TGFBR2 and USP15. a Binding sites of miR-30c-2-3p in TGFBR2-3′-UTR and USP15-3′-UTR. b Relative luciferase activities in indicated cells transfected with miR-30c-2-3p mimic and luciferase reporters containing TGFBR2-3’UTR or USP15-3′UTR (each bar represents the mean ± SD derived from three independent experiments, two-tailed Student’s t test. *P < 0.05; ns, not significant). c The reporters containing TGFBR2-3′-UTR or USP15-3′-UTR were co-transfected with miR-30c-2-3p inhibitor, and luciferase activities were assessed after 48 h (each bar represents the mean ± SD derived from three independent experiments, two-tailed Student’s t test. *P < 0.05; ns, not significant). d Effects of silencing TGFBR2 or USP15 in indicated cells on luciferase activities of the TGF-β responsive reporter (each bar represents the mean ± SD derived from three independent experiments, one-way ANOVA followed by Dunnett’s multiple comparison test. *P < 0.05). e Effects of silencing TGFBR2 or USP15 on cell invasion as measured by Transwell invasion assay in the indicated NSCLC cells (each bar represents the mean ± SD derived from three independent experiments, one-way ANOVA followed by Dunnett’s multiple comparison test. *P < 0.05). f Luciferase assay of reporters for pGL3-TGFBR2-3′-UTR or pGL3-USP15-3′-UTR in indicated cells, co-transfected with MYEOV-cDNA, MYEOV-ATGmut or MYEOV-miRmut (each bar represents the mean ± SD derived from three independent experiments, one-way ANOVA followed by Dunnett’s multiple comparison test. *P < 0.05; ns, not significant). g WB analysis performed for TGFBR2 and USP15 with β-actin as loading control. h Model diagram of proposed MYEOV-mediated regulation of TGF-β signaling in NSCLC