Fig. 4.

Modulation of Trpv1 improves ER–mitochondrial tethering in cells overexpressing Chop. A Immunoblot analysis of Trpv1 in total cell lysates of MLE12 cells untreated or treated with dox for Chop expression for 12 and 24 h. B Relative protein level of Trpv1 were normalized to Gapdh and its level in −dox cells was set as one. C MLE12 cells were either left untreated or treated with dox for 12 h, followed by treatments with DMSO or CPS at indicated concentrations followed by immunoblots for the indicated proteins. D and E Quantification of Trpv1 (D) and Pacs2 (E) protein expression is shown. Relative protein amounts were normalized to Gapdh and their mean value in respective DMSO-treated controls was set as one. F Representative fluorescence images following proximity ligation assay with antibodies against calnexin and VDAC1, and cells treated with CPS at the indicated concentrations or DMSO upon Chop induction are shown, scale bar = 60 µm. G Fluorescence intensity was quantified using ImageJ, its intensity in −dox, DMSO-treated cells was set to one. ‘n’ of three independent experiments were performed and statistical significance is indicated as: *p ≤ 0.05, ***p ≤ 0.01, ***p ≤ 0.001