Research Paper Volume 11, Issue 24 pp 11988—12001

CircRNA AFF4 promotes osteoblast cells proliferation and inhibits apoptosis via the Mir-7223-5p/PIK3R1 axis

Figure 3. miR-7223-5p regulates cells function via targeting FGFR3. (A) Predicted targets of miR-7223-5p. (B) PIK3R1 levels in calluses during fracture healing measured by qRT-PCR. (C) PIK3R1 expression in calluses on days 14 and 21 post-fracture after PBS and agomiR-7223-5p injection. (D, E) PIK3R1 expression measured by western blot and qRT-PCR after transfecting cells with agomiR-7223-5p or antagomiR-7223-5p. (F) The predicted target site of miR-7223-5p on PIK3R1 and luciferase activities. (G, H) CCK8 and Edu staining showing MC3T3-E1 proliferation after transfection with siPIK3R1 or antagomiR-7223-5p+siPIK3R1. (I) Cyclin D1 and cyclin D3 levels in cells measured by Western blot. (J) Cell apoptosis (Q2+Q3) assessed by flow cytometry 24 h after transfection with si-PI3KR1. Q1: dead cell; Q2: later apoptosis; Q3: early apoptosis; Q4: living cells. (K) Apoptosis-related proteins were detected by Western blot.