Fig 2: Effects of telmisartan on Cnx 43, KCNQ1 and Bcl-2 expression in acute myocardial I/R injury. A, Representative images for immunochemical detection of Cnx43 and KCNQ1 levels. B, Bar graph showing Cnx43 and KCNQ1 expressed as percentage of positive stained area/total area. C, Bcl-2 protein levels detected by ELISA. SHAM, thoracotomy only; I/R, ischaemia/reperfusion; veh, 1% methylcellulose 4 mL/kg/d; Tel, telmisartan 12 mg/kg. Results are expressed as mean ± SEM of five observations. **P < .01 vs SHAM; °P < .05 and °°P < .01 vs I/R veh. Scale bar = 50 µmol/L; 400× magnification

Fig 3: (A) Representative immunocytochemistries of H9c2 cells cultured in normal glucose (Normal glu) or 33 mM glucose (High glu) and labelled with KCNQ1 antibody. Cells were exposed to DMSO 0.05 µM and BF-5 m 0.01–0.025–0.05 µM. Representative microscopic fields for each treatment are shown and accordingly, the percentage of KCNQ1-positive cells is represented in the graph. (B) The results are expressed as mean ± S.E.M. of the percentages of positive cell / total cell counted in each analyzed field for each treatment. **P < 0.01 Normal glu; °°P < 0.01 vs High glu. 40× magnification.

Fig 4: Representative merged images of MitoTracker labeling of mitochondria (red), immunofluorescence with KCNQ1 antibody (green) and counterstaining of cell nuclei with Hoechst 33258 (blue) in H9c2 cells cultured in normal glucose (Normal glu) or 33 mM glucose (High glu) and exposed to DMSO 0.05 µM and BF-5m 0.01–0.025–0.05 µMMerged images did not demonstrate a significant overlap between mitochondria and KCNQ1 in H9c2 exposed to normal or high glucose medium and receiving DMSO 0.1 µM or BF-5 m 0.01–0.025–0.05 µM pretreatment. 40× magnification.

Fig 5: (A) qRT-PCR assay of miR-1 expression levels expressed as arbitrary units (A.U.) in H9c2 cultured in normal medium (Normal glu) or high glucose medium (High glu) and exposed to DMSO 0.05 µM and BF-5m 0.01–0.025–0.05 µM. H9c2 grown in high glucose medium showed an up-regulation of miR-1 levels compared to normal glu. H9c2 cultured in glucose 33 mM (High glu) and pretreated with BF-5m 0.01–0.025–0.05 µM showed a significant down-regulation of miR-1 expression levels compared to High glu. (B, C, D) Western blotting analysis of KCNE1 (B, D) and KCNQ1 (C, D) protein levels in H9c2 cells exposed to normal glucose (Normal glu) or high glucose (High glu) and exposed to DMSO 0.05 µM and BF-5m 0.01–0.025–0.05 µM. High glucose significantly decreased KCNE1 and KCNQ1 protein levels. Treatment with BF-5m 0.01–0.025–0.05 µM restored the KCNE1 and KCNQ1 levels. The results are reported as the mean ± S.E.M. of n = 3 treatments. **P < 0.01 vs Normal glu, °P < 0.05 vs High glu and °°P < 0.01 vs High glu.