Fig. 3.

PHF8 facilitates HER2 signalling through its tumour-promoter activity. a. The proliferation of MCF10A cells overexpressing HER2, with PHF8 knockdown, or both was assessed using the MTT assay. Data are presented as mean ± SD of three independent experiments. **p ≤ 0.01. b. Western blotting analysis of the expression of HER2, PHF8, and other indicated proteins shown in panel a in MCF10A cells. The data represent three independent experiments. A and B, Mock: overexpression control; HER2: HER2 overexpression; shNC: scrambled shRNA; shPHF8: PHF8 shRNAs. c. PHF8 knockdown counteracted the activity of most pathways induced by HER2 overexpression. Gene set Enrichment Analysis (GSEA) of Hallmark pathways in HER2-overexpressing cells vs. control cells compared with HER2-overexpressing cells vs. PHF8 knocking down. The p values are coloured; normalised enrichment scores (NES) are shown on the x-axis. d. Heat map of RNA sequencing Z-score results of 298 PHF8 differentially regulated genes (DRG) that were significantly regulated by HER2 overexpression. e. Heat map of the Z-scores of 60 PHF8-associated DRGs contributing most to enriched pathways. f. Protein–protein association networks of 60 PHF8-associated DRGs were analysed using STRING. Edges represent protein-protein associations such as direct interactions, gene neighbourhood, co-expression, and co-occurrence.