Fig. 5

Circ_001422 sponges miR-195-5p. a. A RIP assay was performed to assess the circ_001422 level in 143B cells transfected with Ago2 overexpression vectors or control vectors. b. Schematic illustration showing the overlapping target miRNAs of circ_001422 predicted by TargetScan, miRanda, starBase, RNAhybrid and RNA22. c. Lysates prepared from OS cells transfected with circ_001422 overexpression vectors or control vectors were subjected to an RNA pulldown assay, and the pulldown efficiency was confirmed by qRT-PCR. d. The relative quantification (RQ) values of 4 candidate miRNAs in OS cell lysates were examined by qRT-PCR. e. The putative binding sites between miR-195-5p and circ_001422. f. Luciferase activity was assessed in OS cells after cotransfection with circ_001422-WT or circ_001422-MUT and the miR-195-5p mimic or NC mimic. g-i. An anti-Ago2 RIP assay was conducted in OS cells after transfection with the miR-195-5p mimic or NC mimic prior to western blot and qRT-PCR analyses to measure the expression levels of Ago2, circ_001422 and miR-195-5p. j. The RQ value of miR-195-5p was determined by qRT-PCR in 55 paired OS tissues and adjacent noncancerous tissues. k. Pearson correlation analysis of circ_001422 and miR-195-5p expression in the 55 OS tissues. l. RQ value of miR-195-5p in OS cells and normal osteoblasts. n.s., not significant; ^*P < 0.05; ^**P < 0.01; ^***P < 0.001