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. 2024 Aug 26;15(8):621. doi: 10.1038/s41419-024-06996-w

Fig. 7. BRG1 suppresses PPP2R1A expression by occupying its transcriptional activation site, thereby activating PI3K/AKT signaling.

Fig. 7

A ChIP-Seq was performed on SUP-B15 (ALL_1) and Nalm-6 (ALL_2) cells, and Venn diagram showing the gene overlap analysis between the ChIP-seq data of the two cell lines. B KEGG analysis was performed on the fitted gene clusters, in which the PI3K–AKT signaling pathway was significantly affected, and genes involved in the regulation of the pathway were categorised and analysed, with the grey section representing genes related to extracellular components, the blue section representing genes localised in the cell membrane, and the red section representing genes localised in the cytoplasm. C The effect of BRG1 silencing on the products encoded by genes in the cytoplasm in plot B was analysed in the proteomic data, which showed that the expression of PPP2R1A and PPP2R1B was elevated and the expression of CDK6 was decreased. D Schematic diagram of the regulation of PP2A and PI3K–AKT signaling pathway by BRG1. Given that the results of ChIP-Seq showed that the site of binding of BRG1 to the PPP2R1B is on the intron and the binding site to PPP2R1A is on the promoter, (E) the correlation between PPP2R1A and BRG1 (SMARCA4) was analysed at the protein level. F The expression of PPP2R1A in three cell lines with differential expression of BRG1 was evaluated via western blotting. G Grey value analysis of the protein expression of plot F. # represents statistical differences in BRG1 expression between groups, & represents statistical differences in PPP2R1A expression between groups. H The mRNA expression of PPP2R1A in three cell lines with differential expression of BRG1 was detected by qRT-PCR. I ChIP-seq profiling showed the ChIP-seq signal for BRG1 at the genomic loci of PPP2R1A in ALL_1 (-log10(p-value) = 7.13) and ALL_2 (-log10(p-value) = 3.2). (Red marker indicates PPP2R1A promoter region: Chr19:52188052-52190052). J ChIP-qPCR was used to detect the binding of BRG1 to the PPP2R1A promoter in SUP-B15 and Nalm-6 cells. A dual luciferase plasmid overexpressing PPP2R1A was constructed and transfected with it into BRG1-overexpressing RS4:11 cells (LV-BRG1) and control cells (LV-Vector). K Histogram showing relative fluorescence intensity of groups. (*, & and #, P < 0.05; **P < 0.01; ***P < 0.001; -log10(p-value)>1.3, P < 0.05; ns not significant).