Research Paper Volume 13, Issue 7 pp 10584—10602

Suppression of choroidal neovascularization by silencing of long non-coding RNA IPW

LncRNA-IPW regulates the expression of genes located in DLK1-DIO3 locus. (A) RF/6A cells were transfected with scrambled (Scr) siRNA, lncRNA-IPW siRNA, or left untreated (Ctrl) for 24 h. qRT-PCRs were performed to detect the expression of miR-370, miR-408, or MEG3 (n = 4; *P B) The 3’-end biotinylated miRNA duplexes were transfected into RF/6A cells. After streptavidin capture, the levels of IPW and GAPDH in the input and bound fractions were detected using qRT-PCRs. The relative immunoprecipitate (IP)/input ratios were plotted. (C) qRT-PCRs were performed to detect the expression of miR-370 in the RPE/choroid complex of C57BL/6J mice on day 3, day 7, and day 14 after laser photocoagulation (n = 6 animal per group; *P U test). (D) RF/6A cells were exposed to 200 μM CoCl2 for the indicated time. qRT-PCRs were performed to detect the expression of miR-370 (n = 4; *P t-test).

Figure 5. LncRNA-IPW regulates the expression of genes located in DLK1-DIO3 locus. (A) RF/6A cells were transfected with scrambled (Scr) siRNA, lncRNA-IPW siRNA, or left untreated (Ctrl) for 24 h. qRT-PCRs were performed to detect the expression of miR-370, miR-408, or MEG3 (n = 4; *P < 0.05; One-way ANOVA). (B) The 3’-end biotinylated miRNA duplexes were transfected into RF/6A cells. After streptavidin capture, the levels of IPW and GAPDH in the input and bound fractions were detected using qRT-PCRs. The relative immunoprecipitate (IP)/input ratios were plotted. (C) qRT-PCRs were performed to detect the expression of miR-370 in the RPE/choroid complex of C57BL/6J mice on day 3, day 7, and day 14 after laser photocoagulation (n = 6 animal per group; *P < 0.05; Mann-Whitney U test). (D) RF/6A cells were exposed to 200 μM CoCl2 for the indicated time. qRT-PCRs were performed to detect the expression of miR-370 (n = 4; *P < 0.05; Student’s t-test).