Fig 2: Coexpression of CHOP and HIF-1a in alveolar epithelial cells of bleomycin-induced lung fibrosis in mice. CHOP (A–C) and HIF-1a (D–F) are co-expressed (serial sections) in hyperplastic alveolar epithelial cells (AECs) in and close to areas of pulmonary remodelling (arrow). No labelling was observed with isotypic controls (G–I). Sections were counterstained with Nuclear Fast Red. BC are higher magnifications of A; EF of D and HI of G; Original magnification: objective X200 (A,D,G), x 400 (B,E,H) and x 1000 (C,F,I). Scale bars represent 50 µm.

Fig 3: Knocking down Nrf2 reversed the miR-144-3p inhibitor-alleviated H/R-induced apoptosis, oxidative stress, mitochondrial damage and endoplasmic reticulum stress in vitro. A–c, F–K, The protein expression level of Nrf2, HO-1, cleaved-caspase3, Bax, Bcl-2, CHOP, GRP78 and Cytochrome C in H/R-injured TCMK cells transfected with miR-144-3p inhibitor or si-Nrf2. Representative bands are shown in A, and the quantitative analysis of protein expression is shown in B, C, F–K (n = 3). D, E, The SOD and MDA levels in cellular supernatants in H/R-injured TCMK cells transfected with miR-144-3p inhibitor or si-Nrf2 (n = 3). All data are expressed as the mean ± SD; data comparisons between multiple groups were performed using one-way analysis of variance (ANOVA) with Tukey's post hoc test. *p < 0.05, **p < 0.01, NS, no significant difference

Fig 4: High glucose and oxidized low-density lipoprotein (oxLDL) synergize in increasing CD36 expression in vascular smooth muscle cells (VSMC). Starved VSMC were cultured for 72 h with M199 medium containing 5 mM (normal glucose, NG) or 22 mM of glucose (high glucose, HG) and treated or not with 50 µg/mL oxLDL for the last 24 h. (A–H) CD36, MCP1, IL6, IL-1ß, and endoplasmic reticulum (ER) stress markers (ATF6, HSPA5, ERN-1 and CHOP) mRNA levels were determined by quantitative real-time PCR analysis (qPCR) and normalized to GAPDH. VSMC from at least four different donors were used. Mean fold change relative to untreated VSMC-NG ± SEM from five independent experiments performed in duplicate are shown. * p = 0.05 vs. 5 mM; # p = 0.05 vs. 22 mM; $ p = 0.05 vs. 5 mM + oxLDL.

Fig 5: CD36 and ER stress markers expression in carotid plaques from subjects with and without diabetes. (A,E,G–J) the mRNA levels of CD36 and endoplasmic reticulum (ER) stress markers were determined in sections of carotids plaques from subjects with and without diabetes by qPCR analysis (n = 15). (B) Immunostaining of CD36 in paraffin sections of carotid plaques (n = 5). (C) Representative blot of CD36 in protein lysates from carotid plaques. (D) Histogram showing circulating levels of 7-KC in plasma from subjects with diabetes (n = 18) and subjects without diabetes (n = 18) determined by LC-MS/MS. (F) Representative blots of ATF6 and CHOP in protein lysates from carotid plaques. Results are expressed as mean ± SEM. * p < 0.05 vs. Subjects without diabetes (NO DIA). ** p < 0.01 vs. NO DIA.