Figure 7: MiR-19a-3p/19b-3p regulates human cardiac fibroblasts autophagy-mediated fibrosis induced by TGF-β1.

(a) Effects of miR-19a-3p/19b-3p and TGF-β1 on cell invasion in human cardiac fibroblasts in vitro. Photographs of the cell invasion through the polycarbonate membrane stained with crystal violet. (b) The inhibitory effect of miR-19a-3p/19b-3p and TGF-β1 on the invasion of the cells was quantified. Data were presented as the mean ± SEM of three separate experiments. *p < 0.05, **p < 0.01, group of pro-transfected with miR-19a-3p/19b-3p mimics and stimulation of TGF-β1 compared with control. (c,d) Human cardiac fibroblasts were pro-transfected with miR-19a-3p/19b-3p and then treated with 10 ng/ml TGF-β1 for 72 h. Cells were then probed by SNLYSO sensor and autolysosomes were analyzed and quantitated by flow cytometer. Results from three independent experiments are shown as means ± SEM. **p < 0.01, group of pro-transfected with miR-19a-3p/19b-3p mimics and stimulation of TGF-β1 compared with control. (e) Confocal fluorescence images of endogenous MAP-LC3 and SNLYSO sensor-labeled compartment in human cardiac fibroblasts. Human cardiac fibroblasts were pro-transfected with miR-19a-3p/19b-3p and then treated with 10 ng/ml TGF-β1. MAP-LC3 (green) was labeled with primary anti-MAP-LC3 polyclonal antibody. Goat anti-rabbit IgG/FITC were used as secondary antibody. Cells were simultaneously imaged in the presence of SNLYSO sensor (red) to visualize autophagy. Nuclei (blue) were labeled by DAPI. Confocal microscopy images were obtained. Bar = 30 μm.