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. 2020 Oct 22;26:100335. doi: 10.1016/j.jbo.2020.100335

Fig. 4.

Fig. 4

FOXD3-AS1 enhanced ZCCHC3 expression in OS by absorbing miR-296-5p. (A) ZCCHC3 was screened out to be the target gene of miR-296-5p through PITA, TargetScan, PicTar, microT and RNA22 programs in starBase. (B) ZCCHC3 expression was detected via RT-qPCR in 52 OS tissues and paired non-tumor tissues. Student’s t test. (C) RT-qPCR detected ZCCHC3 expression in OB3 cells and OS cells. One-way ANOVA. (D) RT-qPCR detected miR-296-5p expression in MG-63 and HOS cells after transfection with NC inhibitor or miR-296-5p inhibitor. Student’s t test. (E) RT-qPCR and western blot analyses measured the mRNA and protein levels of ZCCHC3 in different groups. One-way ANOVA. (F) The binding site between miR-296-5p and ZCCHC3 was predicted by starBase. (G) The luciferase activity of ZCCHC3 3′UTR-WT and ZCCHC3 3′UTR-Mut in different groups was determined by luciferase reporter assay. Student’s t test. (H) RIP assay detected the relative enrichment of FOXD3-AS1, miR-296-5p and ZCCHC3 in anti-Ago2 and anti-IgG groups. Student’s t test. **P < 0.01.