Fig 2: Analysis of the effects of treatment with IL-1Ra in HAECs exposed to d-flow. (A) Results of the western blotting of GRP78, IL-1β, IRAK2, CHOP, caspase 12 and GAPDH in HAECs treated with IL-1Ra and/or d-flow. The relative expressions of GRP78 (white columns), IL-1β (light gray columns), IRAK2 (dark gray columns) and CHOP (black columns) were normalized to GAPDH. (B) Images were captured of the terminal deoxynucleotidyl transferase dUTP nick end labeling assay of HAECs treated with IL-1Ra and/or d-flow. The detected apoptotic percentages of HAECs treated by IL-1Ra and/or d-flow were quantified. Magnification, ×200. *P<0.05 vs. Ctrl; #P<0.05 vs. Ctrl+IL-1Ra; &P<0.05 vs. d-flow. IL-1Ra, interleukin-1 receptor antagonist protein; GRP78, 78 kDa glucose-regulated protein; CHOP, DNA damage-inducible transcript 3 protein; d-flow, disturbed blood flow; HAECs, human aortic endothelial cells; Ctrl, control; IRAK2, interleukin-1 receptor associated kinase 2; IL-1β, interleukin-1β.

Fig 3: Effect of BAPTA-AM on Pb-induced protein levels of ER stress markers in rPT cellsCells were pretreated with 10 µM BAPTA-AM for 30 min, then exposed to 0.5 µM Pb for another 12 h to measure the protein levels of GRP78 (A), GRP94 (B), CRT (C), CHOP (D) and caspase-12 (E). Upper panel representative western blot image; lower panel quantitative analysis (mean ± SEM, n = 4). *P < 0.05, **P < 0.01.

Fig 4: Effects of TGF-β 1 on the expression of FN, COL I, GRP78s and CHOP in RMCs. (A) RMCs were stimulated with TGF-β 1 (5 ng/ml) and gene expression of FN, COL I, GRP78 and CHOP were assessed after 6, 12 and 24 h by RT-PCR. (B) Western blot analysis and grayscale analysis for FN, COL I, GRP78 and CHOP. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001; ns, P>0.05 (not significant). RMCs, rat renal mesangial cells; CHOP; C/EBP homologous protein. FN, fibronectin; COL I, collagen I; GRP78, glucose-regulated protein 78; CHOP, C/EBP homologous protein; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; TGF, transforming growth factor.

Fig 5: Knockdown of glucose-regulated protein 78 (GRP78) reduced tumor growth in gastric cancer xenografts. (A) Schematic of gastric cancer xenografts between MKN45/ctrl and MKN45/shGRP78 groups. (B) The curves of tumor growth in mice between MKN45/ctrl and MKN45/shGRP78 groups. (C) Representative images of hematoxylin and eosin and immunohistochemical (IHC) staining of ki-67 between MKN45/ctrl and MKN45/shGRP78 groups. (D) Representative IHC analysis of GRP78 in MKN45/ctrl and MKN45/shGRP78 gastric cancer xenografts and relative quantification per intensity of staining scoring. (E) Representative IHC analysis of ki-67 staining in MKN45/ctrl and MKN45/shGRP78 gastric cancer xenografts and relative quantification per intensity of staining scoring. (F) Curves of body weight of mice between MKN45/ctrl and MKN45/shGRP78 groups. (G) Tumor sizes of each group. Data are presented as the mean ± standard error of mean; n = 4 independent experiments, two-tailed Student’s t test: ** p < 0.01, *** p < 0.005.