Fig 1: ARGLU1 enhanced the transcriptional activity of AP1 and YY1 on MLH3, MSH2, MSH3 and MSH6. (a) Venn diagram of ARGLU1 IP-MS result and transcription factors of 4 MMR genes (MLH3, MSH2, MSH3 and MSH6) predicted by JASPAR. (b) Co-IP result of ARGLU1 with YY1 and SP1. (c) ChIP-qPCR results of YY1 and SP1 in HGC27 cells with or without ARGLU1 knock-down. (d) Relative luciferase activity of MLH3 in shARGLU1, siSP1 or siYY1 treated HGC27 and (e) MGC803 cells. (f) Schematic diagram of the molecular mechanism of ARGLU1 enhancing the transcriptional activity of SP1 and YY1 on MMR genes in GC. The data are the means ± SDs of three independent experiments (two-tailed t-test, *, p < 0.05, **, p < 0.01, ***, p < 0.001, NS, not significant).
Fig 2: ARGLU1 increased the transcriptional level of mismatch repair genes in GC cells. (a) GSEA analysis of ARGLU1 in TCGA-STAD cohort. (b) Quantitative RT-PCR analysis of MMR genes mRNA expression levels in four GC cells with ARGLU1 perturbation (#, |FoldChange|>1.5, p < 0.01). (c) ChIP-seq data exhibited peaks of ARGLU1, H3K4me3, H3Ka27ac and POLR2A in the promoter region of MLH3, MSH2, MSH3 and MSH6. (d) Relative luciferase activity of MLH3, MSH2, MSH3 and MSH4 in different treatment groups of MGC803 and HGC27 cells. The data are the means ± SDs of three independent experiments (two-tailed t-test, *, p < 0.05, **, p < 0.01, ***, p < 0.001, NS, not significant).
Fig 3: Mlh3 depletion impairs oocyte and early embryos developmental potential. A Trim-away workflow. B After 16 h of in vitro culture post microinjection, Mlh3 fluorescence intensity was significantly reduced. C-D Decreased rates of maturation, fertilization, and blastocyst formation after Mlh3 depletion. E CNV analysis of 2-cell-stage embryos from both control and Mlh3-depleted groups. F Blastocyst cell numbers in control versus Mlh3-depleted groups. Pb = polar body. CNV=Copy number variation. p < 0.05, p < 0.01. bar = 20 μm
Fig 4: MLH3 expression in ESC in response to DNA damage stress. (A) mRNA levels after 4-HC treatment. (B-C) Protein levels after 4-HC treatment and recovery. (D) Immunofluorescence analysis of MLH3 expression. *p < 0.05, **p < 0.01, ***p < 0.001. bar = 20 μm
Fig 5: in vivo developmental competence of Mlh3-deficient embryos. A A schematic graph for delivery of Mlh3-deficient and control blastocysts into the mouse uterus. B Images of the uterus, fetus and placentas in the two groups at E13.5. Fetus number (C) and abortion rate (D) of the two groups. Crown-rump length (E), placenta length (F), fetus weight (G) and placenta weight (H) of the two groups. *p < 0.05, **p < 0.01, ***p < 0.001
Supplier Page from Abcam for Anti-MLH3 antibody