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. 2020 Jul 2;295(34):12045–12057. doi: 10.1074/jbc.RA120.012565

Figure 5.

Figure 5.

Ambra1, Akap8, Cdk9, and Atf2 coregulate a subset of genes. SCC FAK-WT cells were transfected with siControl, siAmbra1, siAkap8, and siAtf2 (siGENOME pool). RNA was isolated 48 h posttransfection and subjected to gene expression analysis using the mouse nCounter PanCancer Pathways panel. A, Venn diagram of significantly altered genes compared with siControl (p < 0.05, 2-fold difference compared with siControl). B, relative gene expression of 18 genes coregulated by Ambra1, Akap8, and Atf2 knockdown compared with siControl. C, table of the top five enriched signaling pathway gene sets according to KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis of Ambra1-, Akap8-, and Atf2-regulated genes. D, Western blotting showing Atf2 knockdown by siRNA in SCC FAK-WT cells. Anti-GAPDH served as a loading control. E and F, validation of Ambra1-, Akap8-, and Atf2-mediated Angpt1, Tgfb2, Tgfb3, Itga8, and Itgb7 expression changes by qRT-PCR using RNA isolated 48 h posttransfection of SCC FAK-WT cells with siControl, siAmbra1, siAkap8, and siAtf2 (E) as well as siControl and siCdk9 (F). Error bars represent S.D. *, p < 0.01; #, p < 0.05.