Fig. 6

circ6401 acts as a sponge for miR-29b-1-5p and prevents it from inhibiting the translation of RAP1B in WJ-MSCs. a RNA FISH assay for subcellular co-localization of circ6401 and miR-29b-1-5p in WJ-MSCs. Cy3 labeled the circ6401 FISH probe, FITC labeled the miR-29b-1-5p FISH probe, and DAPI stained the cell nuclei. Scale bar, 20 μm. b 3′ UTR sequences containing the predicted “seed region” target site of circ6401 and its mut-sequences were cloned into the psiCHECK2.0 vector to prove whether circ6401 was the direct target gene of miR-29b-1-5p. c Dual-luciferase reporter gene assay to detect interaction between miR-29b-1-5p and circ6401. circ6401-mut, circ6401 3′UTR mutant; circ6401-wt, circ6401 wild type; mimics NC, empty vector control. ^*P < 0.05. d qRT-PCR was used to determine miR-29b-1-5p expression level of WJ-MSCs transfected with lenti-NC/lenti-circ6401. Lenti-NC, lentivirus with empty vector; lenti-circ6401, lentivirus with circ6401 overexpressing vector. ^*P < 0.05. e RAP1B mRNA expression in WJ-MSCs after co-transfection with circ6401 overexpression vectors and miR-29b-1-5p mimics. Data in panel are shown as means ± standard deviation (n = 3). ^**P < 0.001. ^#P < 0.05 compared to lenti/mimics NC group; ^##P < 0.001 compared to lenti/mimics NC group; ^nsP > 0.05 compared to lenti/mimics NC group