Figure 4. AP-2α interacts with HDAC1/2, and HDAC activity is required for targeted methylation in HNSCC cells.

a) ChIP analysis was performed using AP-2α, SP1, HDAC1, and HDAC2 antibodies in SCC22B cells with and without AP-2α downregulation. ChIP eluate was then utilized for quantitative PCR using CDH1, RASGRF2, MLH1, RASSF1A, and PTEN promoter primers. b) Western blot analysis demonstrating increased acetylation following Trichostatin A (TSA) treatment. c) Bisulfite sequencing analysis was performed in SCC22B cells before and after TSA treatment. Solid circles represent methylated CpGs; whereas, open circles represent unmethylated CpG sites. d) Graphical representation of the average methylation percentage in the 3 target genes analyzed. * P-value = 0.0452; ** P-value = 3.77×10⁻⁶.