Fig. 7

Inhibition of HTRA1 promoted the endoplasmic reticulum autophagy degradation of Col1. Western blot (A) and quantitative data (B) showing the trends of col1 protein expression under different Chx treatment time (0 h, 1 h, 2 h) in cardiac fibroblasts transfected with or without HTRA1 siRNA. Representative Western blot (C) and quantitative data (D) showing the change of Col1 protein expression under different treatments including Chx + MG132, Chx + EerI and Chx + CQ in cardiac fibroblasts transfected with HTRA1 siRNA. Cardiac fibroblasts were respectively treated for 0, 4 and 8 h. Representative Western blot (E) and quantitative data (F) showing the change of Col1 protein expression under different CQ treatment time (0 h, 4 h, 8 h) in cardiac fibroblasts transfected with or without HTRA1 siRNA. G Representative western blot and quantitative data showing the protein expression difference of autophagy indicators including Beclin1 and LC3B between negative control (NC) and HTRA1 siRNA or HTRA1 plasmid group. Grey represented the control group, blue represented the HTRA1 siRNA group, and red represented the HTRA1 plasma group. H Representative electron microscopy showing endoplasmic reticulum autophagosomes or autophagic lysosomes in cardiac fibroblasts, human and mouse heart. The scale bar indicates 1um. Orange box represented a typical colocalization field of view. Red arrow refers to endoplasmic reticulum segments. I Live-cardiac fibroblasts imaging transfected with the ssRFP-GFP-KDEL tandem reporter plasmid for 24 h. The scale bar indicates 2 μm. Counting the number of red dots in cells for evaluating the intensity of endoplasmic reticulum autophagy. Twenty cells in every group were counted. J The band intensities of RFP and RFP-GFP were determined, and the normalized RFP:RFP-GFP ratio was presented