Fig. 5

Upregulation of NF2 contributes to the tumor suppressing function of KAT2B. (A) RNA-Seq identified differentially expressed genes in KAT2B stably overexpressed cells when compared to the control cells. The transcriptome profiles of KAT2B stably overexpressed cells and control cells were analyzed by RNA-Seq and subjected to differential gene expression analysis. After filtering the low expression genes (with reads < 200), there were 1402 genes upregulated (> 2-fold) by KAT2B overexpression in HuCCT1 cells, which include various tumor suppressor genes. (B) Aligned RNA sequencing reads visualized through Integrative Genomics Viewer (IGV) presented the expression levels of KAT2B and NF2 in HuCCT1 cells. (C) ChIP-quantitative real-time PCR was used to analyze the enrichment of KAT2B in the promoter region of the NF2 gene (n = 8). Three primer pairs (P#1, #2 and 3) targeting the promoter region of NF2 were used for the assay. Rabbit immunoglobulin G (IgG) was used as negative control. (D) qRT-PCR confirmed that the mRNA levels of NF2 were upregulated by KAT2B overexpression in both SG231 cells and HuCCT1 cells (n = 4). (E) Western blotting assay showed that the protein levels of NF2 were upregulated by KAT2B overexpression in CCA cells. (F) WST-1 assay was used to evaluate the effects of NF2 siRNAs on the proliferation defect caused by KAT2B overexpression in SG231 and HuCCT1 cells (n = 8). **P < 0.01