Fig. 2
UPR pathways are downregulated in EDEM3-overexpressing HepaRG cells. A EDEM3 expression in HepaRGC and HepaRGEDEM3 cells was determined by western blot. Detection of α-tubulin was used as total protein loading control. B Expression of EDEM3 (green) and PDI (red) was determined in HepaRGC and HepaRGEDEM3 cells by incubation with corresponding antibodies followed by immunofluorescence microscopy. Images were analyzed with the AxioVision SE64 Rel. 4.9.1 Software. The cell nuclei were stained with DAPI (blue). The indicated scale bar is 50 µm. C Expression of UPR and ERAD markers in HepaRGC and HepaRGEDEM3 cells was investigated by western blot with corresponding antibodies. Detection of α-tubulin, β-actin or calnexin was used as internal controls for total protein loading. The asterisk indicates a non-specific reactivity of the p-eIF2α antibody. The bands corresponding to the proteins of interest were quantified from three independent experiments by using the ImageJ software and normalized to corresponding internal controls. Statistical analysis of the relative protein expression using the unpaired t-test is shown (*p < 0.05, **p < 0.01)