Figure 2
YY1 directly binds to MMP-14 promoter to increase its transcription. (a) Scheme and sequence of the intact YY1 binding site (WT) and its mutation (Mut) within the MMP-14 promoter-luciferase reporter vectors. (b) Dual-luciferase assay showing the activity (normalized to Mock + WT) of MMP-14 promoter reporter [pGL3-MMP14 (-1246/ + 199)] and its mutant in SGC-7901 and AGS cells stably transfected with empty vector (mock) or YY1 (mean ± SD, n = 5). (c) Dual-luciferase assay indicating the activity (normalized to sh-Scb + WT) of MMP-14 promoter reporter [pGL3-MMP14 (−1246/+199)] and its mutant in MKN-45 and AGS cells stably transfected with scramble shRNA (sh-Scb) or YY1 shRNA (sh-YY1) (mean ± SD, n = 4). (d) ChIP (using YY1 antibody) and qPCR assay showing the enrichment of YY1 (normalized to 20% input DNA) on the MMP-14 promoter in gastric cancer cells, and those stably transfected with mock, YY1, sh-Scb, or sh-YY1 (mean ± SD, n = 5). (e) Western blot indicating the expression of p65, YY1, and MMP-14 in SGC-7901 cells transfected with mock or YY1, and those co-transfected with sh-Scb or sh-p65. (f and g) Dual-luciferase, ChIP (using YY1 antibody) and qPCR assays showing the activity of MMP-14 promoter reporter [pGL3-MMP14 (-1246/ + 199)] and the binding of YY1 to MMP-14 promoter in gastric cancer cells transfected with mock or YY1, and those co-transfected with sh-Scb or sh-p65 (mean ± SD, n = 5). *P < 0.01 vs. mock, sh-Scb, IgG, or mock + sh-Scb. **P < 0.01 vs. WT.
