Fig. 2.

MiR-451a inhibits AKT/mTOR signaling pathway via PDPK1-induced phosphorylation modification. (A) Western blot was applied to measure the expression of AKT/mTOR signaling pathway associated proteins (AKT, p-AKT, mTOR, and p-mTOR) in transfected HOS and 143B cells with miR-451a mimics, taking β-actin as internal reference. (B) AKT/mTOR signaling pathway associated protein levels were evaluated by western blot analysis. (C) Q-PCR detection of PDPK1 expression level in transfected HOS and 143B cells with miR-451a mimics or mimics NC. (D) Co-IP assay was used to detect the interaction between PDPK1 and AKT proteins in osteosarcoma cells. (E) In vitro phosphorylation experiment was carried out to verify Thr308 as PDPK1-specific phosphorylation site in the sequence of AKT. **P < 0.01.