Figure 3
From: NDRG1 inhibition sensitizes osteosarcoma cells to combretastatin A-4 through targeting autophagy
CA-4 and CQ upregulate NDRG1 expression respectively in OS cells. (a) SJSA and MG63.2 cells were treated with CA-4, and the whole-cell lysates were subjected to immunoblotting of NDRG1 and GAPDH. (b) The NDRG1 double bands in (a) were quantified and normalized according GAPDH. (c) SJSA and MG63.2 cells were treated with CA-4, and the total mRNA was extracted. Real-time PCR was performed to determine changes in NDRG1 mRNA. GAPDH was used as a loading control. (d) The NDRG1 promoter-driven luciferase reporter was transfected into MG63.2 cells. The results are presented as NDRG1 promoter activity relative to control (relative NDRG1 promoter activity). (e) SJSA and MG63.2 cells were treated with CQ, and the whole-cell lysates were subjected to immunoblotting of NDRG1 and GAPDH. (f) The NDRG1 double bands in (e) were quantified and normalized according GAPDH. (g) Control and CQ-treated OS cells were exposed to 50 μM CHX, a protein synthesis inhibitor. Cells were harvested at the indicated times (0–8 h) after treatment and analyzed by immunoblotting for NDRG1 and GAPDH. (h) NDRG1 levels in (c) were quantified and normalized to GAPDH levels, and half-life of NDRG1 was determined by regression analysis. The data were presented as mean±S.D (*P<0.05, n=3)
