AKT regulates UGCG expression via epigenomic alterations.
<p>(<b>A</b>) A schematic diagram showing AKT-mediated phosphorylation of DNMT1 leading to hypomethylation of CpG islands that further enhances UGCG expression. (<b>B</b>) Immunoblot showing alteration in phosphorylation of DNMT1 by pan-phospho-ser antibody in MCF-7_RIC...
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2025
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| Summary: | <p>(<b>A</b>) A schematic diagram showing AKT-mediated phosphorylation of DNMT1 leading to hypomethylation of CpG islands that further enhances UGCG expression. (<b>B</b>) Immunoblot showing alteration in phosphorylation of DNMT1 by pan-phospho-ser antibody in MCF-7_RICTOR<sup>SH</sup> and MCF-7_ZFX<sup>OE</sup> cells compared to MCF-7 cells. (<b>C</b>) Immunoblots showing a dose-dependent decrease in pAKT and UGCG expression in MCF-7_ZFX<sup>OE</sup> cells on treatment with AKT inhibitor MK2206. (<b>D</b>) Results from qRT-PCR (mean ± SEM, <i>n</i> = 4) show a decrease in <i>UGCG</i> expression in MCF-7_ZFX<sup>OE</sup> cells on AKT inhibition by MK2206. (<b>E</b>) Immunoblot reveals a decrease in phosphorylation of DNMT1 on treatment of MCF-7_ZFX<sup>OE</sup> cells with AKT inhibitor MK2206. (<b>F</b>, <b>G</b>) Results from qRT-PCR (mean ± SEM, <i>n</i> = 3) (F) and immunoblot (G) show increased <i>UGCG</i> expression in MCF-7_RICTOR<sup>SH</sup> cells on DNMT inhibition by DAC. (<b>H</b>) ChIP-qPCR results (mean ± SEM, <i>n</i> = 3) confirm enhanced binding of ZFX to UGCG promoter in MCF-7_RICTOR<sup>SH</sup> cells on DAC (5 μM) treatment. (<b>I</b>) A schematic representation of pAKT-mediated regulation of histone demethylase KDM5A that regulates UGCG transcription via histone methylation. (<b>J</b>) Immunoblot showing the change in phosphorylation of KDM5A using the pan-phospho-Ser antibody in MCF-7_RICTOR<sup>SH</sup> cells compared to MCF-7 cells. (<b>K</b>, <b>L</b>) Immunoblot (K) and its quantification (mean ± SEM, <i>n</i> = 3) (L) show alterations in KDM5A expression in nuclear and cytoplasmic extracts in MCF-7_RICTOR<sup>SH</sup> cells compared to MCF-7 cells. (<b>M</b>) Immunoblot reveals a decrease in phosphorylation of KDM5A on treatment of MCF-7_ZFX<sup>OE</sup> cells with AKT inhibitor MK2206. (<b>N</b>, <b>O</b>) Results from qRT-PCR (mean ± SEM, <i>n</i> = 3) (<b>N</b>) and immunoblot (O) show increased <i>UGCG</i> expression in MCF-7_RICTOR<sup>SH</sup> cells on KDM5A inhibition. (P) ChIP-qPCR results (mean ± SEM, <i>n</i> = 3) show a reduced H3K4Me3 mark on UGCG promoter in MCF-7_RICTOR<sup>SH</sup> cells that increases on treatment with KDOAM-25 inhibitor (30 μM). Data among two groups were analyzed using an unpaired Student <i><i>t</i></i> test and among multiple groups using One-way ANOVA. <i><i>p</i></i>-value: *<i><i>p</i></i> < 0.05, **<i><i>p</i></i> < 0.01, ***<i><i>p</i></i> < 0.0005, ****<i><i>p</i></i> < 0.0001. Numerical data can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003362#pbio.3003362.s015" target="_blank">S4 Data</a>.</p> |
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