Figures & data
Figure 1. OGD/R stimulation-induced damages in H9c2 cells
After OGD/R induction, (A) cell viability was declined and (B) apoptotic cells were increased; (C-D) the production of Bax, Cleaved-PARP, and Cleaved-caspase 3 was promoted. OGD/R, oxygen-glucose deprivation/reperfusion; PARP, poly ADP-ribose polymerase. **P < 0.01, or ***P < 0.001.
Figure 2. OGD/R deactivated ERK and mTOR pathways
After OGD/R induction, (A) the generation of ERK and mTOR and (B) the p/t-ERK as well as (C) the production of p-p70S6K, p-mTOR, (D) p/t-p70S6K and p/t-mTOR were declined. OGD/R, oxygen-glucose deprivation/reperfusion; ERK, extracellular regulated protein kinases; mTOR, mammalian target of rapamycin. *P < 0.05, **P < 0.01 or ***P < 0.001.
Figure 3. Overexpressed circRNA-DENND4C and under-expressed circRNA-DENND4C in OGD/R-induced H9c2 cells
After the OGD/R stimulation, (A) circRNA-DENND4C was overexpressed; (B) after transfection, the expression of circRNA-DENND4C was enhanced and/or decreased by circRNA-DENND4C overexpressing plasmid and/or si-circRNA-DENND4C; after transfection and OGD/R stimulating, (C) viability was positively controlled by circRNA-DENND4C; (E and F) the apoptotic cells and (D) the expression of Bax, Cleaved-PARP, and Cleaved-caspase 3 were negatively controlled by circRNA-DENND4C. OGD/R, oxygen-glucose deprivation/reperfusion; PARP, poly ADP-ribose polymerase; circRNA-DENND4C, circular RNA-differentially expressed in normal cells and neoplasia domain containing 4 C. *P < 0.05, **P < 0.01 or ***P < 0.001.
Figure 4. CircRNA-DENND4C directly targeted miR-320 in H9c2 cells
(A) After the transfection of circRNA-DENND4C overexpressing plasmid and si-circRNA-DENND4C, miR-320 was negatively controlled by circRNA-DENND4C, and (B) dual-luciferase reporter assay confirmed that the miR-320 was targeted by circRNA-DENND4C; (C) after the transfection of miR-320 mimic, the RNA level of miR-320 was increased. CircRNA-DENND4C, circular RNA-differentially expressed in normal cells and neoplasia domain containing 4 C; OGD/R, oxygen-glucose deprivation/reperfusion; miR-320, microRNA-320; NC, negative control. *P < 0.05, **P < 0.01 or ***P < 0.001.
Figure 5. miR-320 mimic impeded H9c2 cell’s growth by facilitating apoptosis and hindering viability
After the co-transfection of circRNA-DENND4C overexpressing plasmid and miR-320 mimic as well as OGD/R induction, (A and E) cell viability was inhibited by miR-320 mimic; (B and F) apoptotic cells, and (C-D and G-H) the generation of Bax, Cleaved-PARP/caspase 3 were elevated by miR-320 mimic. CircRNA-DENND4C, circular RNA-differentially expressed in normal cells and neoplasia domain containing 4 C; OGD/R, oxygen-glucose deprivation/reperfusion; PARP, poly ADP-ribose polymerase; miR-320, microRNA-320; NC, negative control. *P < 0.05, **P < 0.05 or ***P < 0.001.
Figure 6. Overexpressed circRNA-DENND4C promoted activation of ERK and mTOR pathway by down-regulating miR-320
After co-transfection of circRNA-DENND4C overexpressing plasmid and miR-320 mimic as well as OGD/R induction (A) the expression of ERK and mTOR, (B) the p/t-ERK, (C) the production of p-p70S6K, p-mTOR and (D) the p/t-p70S6K and p/t-mTOR were elevated by overexpressed circRNA-DENND4C but were suppressed by miR-320 mimic. OGD/R, oxygen-glucose deprivation/reperfusion; miR-320, microRNA-320; circRNA-DENND4C, circular RNA-differentially expressed in normal cells and neoplasia domain containing 4 C; NC, negative control; ERK, extracellular regulated protein kinases; mTOR, mammalian target of rapamycin. *P < 0.05, **P < 0.01 or ***P < 0.001.
