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. 2016 Dec 21;8(6):9079–9092. doi: 10.18632/oncotarget.14071

Figure 4. The effect of RUNX3 knockdown on the expression of PTHrP in OSCC cells and RANKL expression in osteoblasts.

Figure 4

(A) shCTRL or shRUNX3 Ca9.22 cells (1 × 105 cells/dish) were treated with 10 ng/ml TGF-β for 24 h. The expression level of PTHrP was examined in the total lysates with a Western blot analysis. GAPDH served as a loading control. (B) shCTRL or shRUNX3 Ca9.22 cells (1 × 103 cells/well) were treated with 10 ng/ml TGF-β for 24 h. PTHrP levels in the conditioned media were analyzed with a ELISA kit. The results are expressed as the mean ± SE. *P < 0.05 versus shCTRL cells without TGF-β. (C) hFOB1.19 osteoblastic cells (1 × 106 cells/dish) were incubated with the respective shCTRL or shRUNX3 Ca9.22 cell-derived conditioned media for 6 h. The expression levels of RANKL and OPG in osteoblastic cells were determined with a Western blot analysis. GAPDH served as a loading control. (D) hFOB1.19 cells (1 × 104 cells/well) were incubated for 24 h with or without conditioned media derived from shCTRL or shRUNX3 Ca9.22 cells. The culture media from hFOB1.19 cells were analyzed for the secreted levels of RANKL and OPG with ELISA kits. The results are expressed as the mean ± SE. *P < 0.05 versus control (without conditioned media), #P < 0.05 versus conditioned media derived from untreated shCTRL cells.