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. 2017 Aug 12;8(47):83088–83099. doi: 10.18632/oncotarget.20236

Figure 5. Transcription factors NF-κB and AP-1 are essential for CAIX-dependent MMP-9 expression.

Figure 5

(A) The reporter constructs of full-length or mutant (the mutation of the NF-κB or AP-1 binding elements) MMP-9 promoter were cloned in the pGL3 luciferase reporter vector. The luciferase reporter assay was used to analyze the effects of the mutation of the NF-κB or AP-1 binding elements on the MMP-9 promoter activity of cell extracts. The results are expressed as mean ± SD of three replications. *p < 0.05 versus vector control; #p < 0.05 versus CAIX expression. The data are presented from three independent experiments. (B) The levels of NF-κB, c-Jun, and c-Fos in SCC-9 cells with or without CAIX expression were determined through Western blot analysis of the nuclear fraction. C23 was used as a loading control. (C) ChIP assay was performed to determine the binding of NF-κB or AP-1 to the MMP-9 promoter. IgG was the negative control, and it was input as the positive control. (D) Proposed mechanism by which CAIX induces oral cancer cell motility.