Fig. 2: TGF-β1 induced autophagy activation in HPMCs, which was confirmed using transmission electron microscopy (TEM), an autophagy flux assay, and western blotting analysis. | Cell Death & Disease

Fig. 2: TGF-β1 induced autophagy activation in HPMCs, which was confirmed using transmission electron microscopy (TEM), an autophagy flux assay, and western blotting analysis.

From: Autophagy caused by oxidative stress promotes TGF-β1-induced epithelial-to-mesenchymal transition in human peritoneal mesothelial cells

Fig. 2

A Representative TEM images of the autophagic morphology. Autophagosomes were frequently observed after TGF-β1 treatment (2 and 5 ng/mL) in HPMCs. The red boxed portion is shown at high magnification on the right. The red arrow indicates the autophagosomes. B TGF-β1-induced autophagy was evaluated by staining using a Cyto-ID Autophagy Detection Kit. Rapamycin (0.5 μM; Rap) was used as a positive control. The stained cells were observed and photographed under fluorescence microscopy (blue, nucleus/ Hoechst 33342; green, autophagosomes/ Cyto-ID). C The intensity of the Cyto-ID green was quantified using a plate reader. D TGF-β1 (2 and 5 ng/mL) increased the mRNA expression of Beclin 1, LC3B, and ATG5, and decreased the mRNA expression of p62. (E, F) TGF-β1 (2 and 5 ng/mL) increased the protein levels of Beclin 1, LC3B, and ATG5, and decreased the protein levels of p62. The results were calculated as values relative to the control. Data are presented as mean ± standard error (SE). n = 4 per group. *P < 0.05 vs. control; **P < 0.01 vs. control; and ***P < 0.001 vs. control.

Back to article page