Research Paper Volume 13, Issue 8 pp 11315—11335

lncRNA C2dat2 facilitates autophagy and apoptosis via the miR-30d-5p/DDIT4/mTOR axis in cerebral ischemia-reperfusion injury

C2dat2 bound to miR-30d-5p and reduced miR-30d-5p expression. (A) Binding region between the miR-30 family and C2dat2, which was predicted by RNAhybrid software. (B and C) Dual-luciferase reporter assays demonstrated that C2dat2 directly targeted miR-30d-5p. (D and E) The luciferase reporter vector carrying C2dat2-WT or the empty vector was cotransfected with the miR-30d-5p mimic or mimic-NC or miR-30d-5p inhibitor or inhibitor-NC into N2a cells as indicated. The relative luciferase activity was detected 48 h after transfection. **P F) The relative expression level of miR-30d-5p was detected via RT-qPCR after C2dat2 knockdown or overexpression. *P **P ##P n = 3 per group). (G) CaMK2d protein expression was measured in N2a cells after OGD/R. (H and I) The relative expression level of CaMK2d was detected via RT-qPCR after C2dat2 knockdown or overexpression at the normal and OGD/R states.

Figure 6. C2dat2 bound to miR-30d-5p and reduced miR-30d-5p expression. (A) Binding region between the miR-30 family and C2dat2, which was predicted by RNAhybrid software. (B and C) Dual-luciferase reporter assays demonstrated that C2dat2 directly targeted miR-30d-5p. (D and E) The luciferase reporter vector carrying C2dat2-WT or the empty vector was cotransfected with the miR-30d-5p mimic or mimic-NC or miR-30d-5p inhibitor or inhibitor-NC into N2a cells as indicated. The relative luciferase activity was detected 48 h after transfection. **P < 0.01 vs. C2dat2-WT. (F) The relative expression level of miR-30d-5p was detected via RT-qPCR after C2dat2 knockdown or overexpression. *P < 0.05; **P < 0.01 vs. normoxia; ##P < 0.05 vs. si-NT or pcDNA-NC, subjected to the same OGD/R (n = 3 per group). (G) CaMK2d protein expression was measured in N2a cells after OGD/R. (H and I) The relative expression level of CaMK2d was detected via RT-qPCR after C2dat2 knockdown or overexpression at the normal and OGD/R states.