Figure 2. RNA sequencing (RNAseq) analysis of ERK1-silenced T47D cells. (A) RT-qPCR and western blotting showed that ERK1 siRNA decreased ERK1 mRNA and protein expression in T47D cells. (B) Volcano plot showed that ERK1 silencing increased a set of 780 genes in abundance of log2FC ≥ 1, while a set of 188 genes decreased in abundance of log2FC ≤ -1, based on transcriptome sequencing of control group and ERK1 siRNA group. (C) KEGG pathway enrichment analysis of differentially expressed genes in ERK1 siRNA group compared to control group. (D) Validation of identified genes in “Hippo signaling pathway”, including YAP1, LEF1, TCF7L1, TCF7L2, AMOT, BTRC, BMP4 and PPP2R1B. (E) Gene Ontology annotation analysis of the significantly enriched cellular component (P < 0.05). (F) Gene Ontology annotation analysis of the top 20 significantly enriched Molecular Function (P < 0.05) that changes in gene expression. (G) Gene Ontology annotation analysis of the top 20 significantly enriched Biological Process (P < 0.05) that changes in gene expression. H. Validation of genes in “negative regulation of cell proliferation”, including GPER1, NR2E3, GATA3, BCL6, ERBB4, NKX3-1 and CDKN1A. **, p<0.01; ***, p<0.001.