Transcription factor EB (TFEB) improves ventricular remodeling after myocardial infarction by inhibiting Wnt/β-catenin signaling pathway

Figure 1. Extent of MI, myocardial hypertrophy and systolic function of heart. (A) Procedure of mice MI modeling after injecting with AAV9. (B) Echocardiography of the mice and the qualified EF ratios in each group. (C) MI areas display in the Masson-stained mouse hearts. Scale bars represent one mm. (D) Ratios of MI areas to the ventricular cavity. (E) Myocyte areas four weeks after MI modeling in different groups (qualified from the WGAs staining). Scale bars represent 50 µm. (F) Survival curves of mice in each group (*: p < 0.05, **: p < 0.01).

Figure 2. TFEB affected the differentiation of fibroblasts and extracellular matrix synthesis and transformation in vivo. MI areas stained by Sirius red four weeks after MI. Collagen I was stained into orange and Collagen III was stained into green. Scale bars represent 10 µm. The expression levels of Collagen I and Collagen III were quantified and presented as mean ± standard deviation. (*: p < 0.05, **: p < 0.01, ****: p < 0.0001).

Figure 3. TFEB Affected CFs in Fibrosis model in vitro. (A) Effects of TGF- β1 of different concentrations and incubation time on proliferation of CFs of rats were measured by CCK-8 assay. (B) The expression of α-SMA was detected using western blotting. (C) Cell viability assessed by CCK-8 assay after rat CFs were treated with increasing concentration of TGF- β1 for 12 h. (D) Cell migration was evaluated by the Transwell assay for the unstimulated CFs (control) and the CFs co-incubated with TGF- β1 (5 ng/mL) for 12 h. (E) Cell migration was evaluated by wound healing assay for the unstimulated CFs (control) and the CFs co-incubated with TGF- β1 (5 ng/mL) for 24 h. Scale bars represent 500 µm. (F) The expression difference of Collagen I in CFs was detected using western blot (*: p < .05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001, ##: p < 0.01).

Figure 4. TFEB relocated to nuclear and made connection with Wnt pathway by β-catenin-TCF/LEF1 complex. (A) Immunofluorescence staining of TFEB (GFP) and β-catenin (red) of the CFs co-incubated with TGF- β1 (5 ng/mL) for 12 h. The interaction between TFEB and β-catenin was showed using high resolution confocal microscopy analysis s. PNU-74654 (100 ng/mL) was used to for 12 h to inhibit β-catenin-TCF/LEF1 complex (40X). Scale bars represent 100 µm. (B) Immunofluorescence staining of TFEB (GFP) and β-catenin (red) of the CFs co-incubated with TGF- β1 (5 ng/mL) for 12 h. Si-TFEB was used to inhibit TFEB (40X). Scale bars represent 100 µm. (C) The expression of α-SMA was detected using western blotting. PNU-74654 (100 ng/mL) was used to for 12 h to inhibit β-catenin-TCF/LEF1 complex. (D) Cell viability assessed by CCK-8 assay after rat CFs were treated with TGF- β1 (5 ng/mL) for 12 h. PNU-74654 (100 ng/mL) was used to for 12 h to inhibit β-catenin-TCF/LEF1 complex. (E) Cell migration was evaluated by the Transwell assay for the CFs co-incubated with TGF- β1 (5 ng/mL) for 12 h. PNU-74654 (100 ng/mL) was used to for 12 h to inhibit β-catenin-TCF/LEF1 complex. (F) Cell migration was evaluated by…

Figure 5. Inhibition effect of TFEB in fibrosis could be mediated by the formation TFEB- β-catenin-TCF/LEF1 complex.