1. (A, B) Representative TEM images from the skin ...
(A, B) Representative TEM images from the skin of healthy volunteers (panels A, A′) and SSc patients (panels B, B′); arrows indicate autolysosomes. The expressions of collagen I, (C) CTGF and (D) LC3 in fibroblasts from healthy volunteers and SSc patients were determined by western blotting analysis. Values are presented as mean (s.d.) (n ≥ 3). *P < 0.05, **P < NC, normal controls (healthy volunteers) TEM: transmission electron microscopy.
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2. HIF-1α and autophagy-related markers P62 and LC3 (A, ...
HIF-1α and autophagy-related markers P62 and LC3 (A, B), together with the protein levels of collagen I and CTGF (C) in fibroblasts were determined by western blot analysis. Levels of HIF-1α, P62 and LC3 (D, E), VE-cadherin, CD31, vimentin and α-SMA (F) in HUVECs were also evaluated by western blot analysis. Values are presented as mean (s.d.) (n ≥ 3). *P < 0.05, **P < 0.01.
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3. (A) LC3 ...
(A) LC3 immunofluorescence images. Cells were exposed to 50 nM (fibroblasts) or 5 nM (HUVECs) bafilomycin A1 and 25 μM (fibroblasts) or 2.5 μM (HUVECs) 2-ME. The staining of antibodies against LC3 (red or green) are for autophagosomes. (B, C) Representative confocal images of fluorescent LC3 highlights cells exposed to 2-ME after being infected with adenovirus GFP-mRFP-LC3. Yellow highlights in merged images represent autophagosomes, while autolysosomes were visualized as red puncta in merged images. At least 20 cells were scored in each experiment. The staining of DAPI (blue) is for the nucleus. Values are presented as mean (s.d.). *P < 0.05, **P < 0.01.
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4. Fig. 4 Autophagy inhibitor bafilomycin A1 and 3-MA reduce hypoxia-induced fibrosis in SSc fibroblasts and endoMT in ...
The protein levels of P62 and LC3 in fibroblasts (A, E) and HUVECs (C, G), collagen I and CTGF in fibroblasts (B, F), VE-cadherin, CD31, vimentin and α-SMA in HUVECs (D, H) were determined by western blot analysis under the hypoxic condition for 24 h or 48 h. Values are presented as mean (s.d.) (n ≥ 3). *P < 0.05, **P < 0.01.
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5. The ...
The protein levels of P62, LC3-II (A), collagen I and CTGF (B) in fibroblasts and P62, LC3-II (C), VE-cadherin, CD31, vimentin and α-SMA (D) in HUVECs were detected by western blot analysis after cells were incubated under the hypoxic condition for 24 h or 48 h. Values are presented as mean (s.d.) (n ≥ 3). *P < 0.05, **P < 0.01.
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6. Fig. 6 The schematic diagram of autophagy involved in this study
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