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. 2015 Dec 29;17(1):35. doi: 10.3390/ijms17010035

Figure 6.

Figure 6

Analysis of the hST3Gal V promoter activity in SD-induced MG-63 cells. The schematic diagrams represent DNA constructs (A) containing various lengths of the wild-type hST3Gal V promoter or constructs (B,C) with the mutant Runx2 sequence in the 5′-flanking region, upstream of a luciferase reporter gene; the transcription start site is designated +1. The pGL3-basic construct, which did not contain a promoter or an enhancer, was used as a negative control. Each construct was transfected into MG-63 cells, with pRL-TK co-transfected as an internal control. The transfected cells were incubated in the presence (open bar) or absence (solid bar) of 10% FBS for 24 h. Relative firefly luciferase activity was measured using the Dual-Luciferase Reporter Assay System, and all firefly activity was normalized to the Renilla luciferase activity derived from pRL-TK. The values represent the means ± SD of three independent experiments with triplicate measurements. ** p < 0.01; *** p < 0.001; (D) PCR amplification in the −432 and −177 region of the hST3Gal V promoter on chromatin immunoprecipitated with either an antibody against Runx2 or control IgG from MG-63 cells grown in the presence or absence of 10% FBS for 24 h. The input (10-fold diluted) represents the positive control.