Research Paper Volume 11, Issue 21 pp 9794—9810

LHX2 promotes malignancy and inhibits autophagy via mTOR in osteosarcoma and is negatively regulated by miR-129-5p

Figure 3. LHX2 knockdown enhances autophagy and inhibits mTOR signaling in vitro. (A) Pearson correlation analysis between LHX2 and autophagy-related genes in 127 OS patients. (B) qRT-PCR analysis of autophagy-related gene expression in LHX2 knockdown cells. *P <0.05, **P<0.01. (C) Representative confocal images of LC3 in lv-GFP-RFP-LC3–infected 143B and MG63 cells. Scale bar: 20 μm (D) LC3 puncta in each cell were counted under 100× magnification. **P<0.01. (E) Autophagy-related protein (LC3B, Beclin1, SQSTM1) levels in lv-control or lv-shLHX2-infected 143B and MG63 cells. (F) 143B and MG63 cells were treated with or without CQ (20 μM) for 12 h. LHX2 and LC3B proteins were detected by western blot. (G) Western blots of p-Akt (Ser473), Akt, p-mTOR (Ser2448), mTOR, p-ULK1 (Ser757), ULK1 in 143B and MG63 cells.