dnmt1 sirna  (Santa Cruz Biotechnology)

Journal: Scientific reports

Article Title: Acetylation-enhanced Sp1 transcriptional activity suppresses Mlph expression.

doi: 10.1038/s41598-025-86282-7

Figure Lengend Snippet: Fig. 2. Effect of DNMT inhibition on melanosome transport. (A) Melanosome images were captured in a bright field, and each image was photographed at a magnification of × 100. (B) The protein expression of Rab27a and Mlph was analyzed using western blot analysis. Melan-a melanocytes were treated 1 or 10 μM 5-aza for indicated times. (C) The protein expression of Rab27a, Mlph, and Myo-Va was analyzed using western blot analysis. Melan-a melanocytes were transfected with 5, 10, or 20 nM of DNMT1-specific siRNA for 72 h.

Article Snippet: DNMT1 siRNA was purchased from Santa Cruz (Santa Cruz, CA, sc-35203). siRNA oligonucleotides including negative control were synthesized by Bioneer (Daejeon, Korea).

Techniques: Inhibition, Expressing, Western Blot, Transfection

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting of the G9a, DNMT1 and UHRF1 epigenetic complex as an effective strategy against pancreatic ductal adenocarcinoma

doi: 10.1186/s13046-024-03268-5

Figure Lengend Snippet: Expression of G9a, DNMT1, and UHRF1 mRNA in human PDAC . A Scoring of G9a, DNMT1 and UHRF1 expression in pancreatic tissues from PDAC patients. Each image shows the representative IHC staining of the three proteins according to each score assigned. B Differential G9a, DNMT1 and UHRF1 expression (3 +) versus (2 + /1 + /0) in tumor tissue and normal pancreatic ducts. C DFS and ( D ) OS in PDAC patients according a combined high expression (3 +) or low expression (2 + /1 + /0) of DNMT1/G9a/UHRF1. *p < 0.05

Article Snippet: Human-specific siRNAs targeting G9a, DNMT1, and UHRF1, along with control siRNA (siC), were sourced from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Expressing, Immunohistochemistry

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting of the G9a, DNMT1 and UHRF1 epigenetic complex as an effective strategy against pancreatic ductal adenocarcinoma

doi: 10.1186/s13046-024-03268-5

Figure Lengend Snippet: Expression and simultaneous targeting of G9a, DNMT1 and UHRF1 in human PDAC cell lines. A G9a, DNMT1 and UHRF1 mRNA levels in PDAC cell lines according to their classification as classical ( n = 9) or quasimesenchymal, QM (basal) ( n = 36) categories extracted from Collisson dataset. B Immunoprecipitation of endogenous G9a in MIA PaCa-2 cells. Immunoprecipitates were probed with anti-G9a, anti-DNMT1 and anti-UHRF1 antibodies. Pre-immune IgG immunoprecipitates and Inputs are shown as controls. C Combination study of the growth inhibitory effects of G9a (UNC0642, UNC) and DNMT1 (AZA) inhibitors in MIA PaCa-2 and PANC-1 cells. D GI 50 values for CM272 in the indicated human and mouse PDAC cell lines. E Effect of CM272 treatment (72 h) on the levels of H3K9me2 in MIA PaCa-2 and PANC-1 cells. H3 total (H3T) levels and isolated histones (Ponceau) are also shown. F Effect of CM272 treatment (72 h) on the global levels of DNA CpG methylation in MIA PaCa-2 and PANC-1 cells. G Colony formation assays in MIA PaCa-2 and PANC-1 cells treated with CM272 at half of their respective GI 50 . *p < 0.05; **p < 0.01

Article Snippet: Human-specific siRNAs targeting G9a, DNMT1, and UHRF1, along with control siRNA (siC), were sourced from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Expressing, Immunoprecipitation, Isolation, CpG Methylation Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting of the G9a, DNMT1 and UHRF1 epigenetic complex as an effective strategy against pancreatic ductal adenocarcinoma

doi: 10.1186/s13046-024-03268-5

Figure Lengend Snippet: The simultaneous inhibition of the epigenetic complex G9a/DNMT1/UHRF1 activates the expression of immune response-related genes in PDAC cells. A qPCR analysis of CCL5, HLA-A, HLA-B and HLA-C expression in MIA PaCa-2 cells pretreated with CM-272 for 24 h and then induced with IFNγ (100U/mL) for another 24 h. B qPCR analysis of CCL5, HLA-A, HLA-B and HLA-C expression in MIA PaCa-2 cells transfected with G9a, DNMT1 or UHRF1-specific siRNAs, specific siRNAs combinations (siG9a + siDNMT1 and siG9a + SiDNMT1 + siUHRF1) and control siRNAs (siC) for 48 h. C qPCR analysis of CCL5, HLA-A, HLA-B and HLA-C expression in MIA PaCa-2 cells treated with UNC, AZA, and the combination (UNC + AZA) at their GI 50 during 72 h. *p < 0.05; **p < 0.01; ***p < 0.001; ns : not significant

Article Snippet: Human-specific siRNAs targeting G9a, DNMT1, and UHRF1, along with control siRNA (siC), were sourced from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Inhibition, Expressing, Transfection, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting of the G9a, DNMT1 and UHRF1 epigenetic complex as an effective strategy against pancreatic ductal adenocarcinoma

doi: 10.1186/s13046-024-03268-5

Figure Lengend Snippet: Overview of the main conclusions of the study. The combined overexpression of DNMT1, G9a, and UHRF1 in PDAC is a strong predictor of poor prognosis. CM272, by targeting this epigenetic complex, shows promising therapeutic potential by inducing apoptosis, reprogramming metabolic pathways, and enhancing immune responses. The combination of CM272 with immunotherapy offers a novel, effective treatment strategy for PDAC

Article Snippet: Human-specific siRNAs targeting G9a, DNMT1, and UHRF1, along with control siRNA (siC), were sourced from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Over Expression

Journal: Molecular Cancer

Article Title: Co-transcriptional R-loops-mediated epigenetic regulation drives growth retardation and docetaxel chemosensitivity enhancement in advanced prostate cancer

doi: 10.1186/s12943-024-01994-0

Figure Lengend Snippet: IGF2BPs upregulate SEMA3F expression via repelling DNMT1. A DNA methylation levels of the SEMA3F gene in TCGA prostate cancer tissues from different groups (blue lines represented the SEMA3F high expression group, purple lines represented the SEMA3F low expression group) were shown by MEPRESS. B Identification of dsDNA specific binding proteins by pull-down. Western blot indicated pulldown of DNMT proteins from PC-3 nuclear extract. C Western blot assay showed the protein levels of SEMA3F in DNMT1 and DNMT3A knockdown compared to control. D Overexpression of IGF2BPs altered global DNA methylation patterns which represented more hypomethylated CpG islands, CGI shelves and shores. E DNA methylation levels of hyper-methylated SEMA3F CpG sites (red square box) decreased in IGF2BPs overexpression teams as compared with control. F ChIP-qPCR analysis showed that IGF2BPs overexpression leads to more DNMT1 binding of SEMA3F promoter. G DNMT1 activities in wild-type and RRM mutant IGF2BPs groups were lower than control and KH1-4 mutants. Data are presented as means ± SD, two-tailed unpaired t-test

Article Snippet: Antibodies used in this study were: anti-Myc tag (ab32, Abcam), anti-IGF2BP1 (ab290736, Abcam), anti-IGF2BP2 (ab128175, Abcam), anti-IGF2BP3 (ab177477, Abcam), anti-RBM15 (10587-1-AP, Proteintech), anti-DNMT1 (ab92314, Abcam), anti-DNMT3A (ab307503, Abcam), anti-DNMT3B(67,259, Cell Signaling Technology), anti-SEMA3F (SAB2107196, Sigma), anti-S9.6 (MABE1095, Sigma), anti-5-mC (ab214727, Abcam), anti-m 6 A (SAB5600251, Sigma), anti-YAP1(phosphor S127, ab76252, Abcam), anti-YAP1(66900-1-Ig, Proteintech), anti-LATS1(66569-1-Ig, Proteintech), anti-LATS2(20276-1-AP, Proteintech), anti-β-Actin(4970, Cell Signaling Technology). siRNAs against DNMT1 used were purchased from Ribobio Co.,Ltd (Guangzhou, China). pcDNA3-based vectors encoding wild-type, RRM domain mutant, KH domain mutant Myc-tagged IGF2BP1, IGF2BP2, IGF2BP3 were produced by Shanghai Yoche Biotechnology Co.,Ltd (Shanghai, China).

Techniques: Expressing, DNA Methylation Assay, Binding Assay, Western Blot, Knockdown, Control, Over Expression, Methylation, ChIP-qPCR, Mutagenesis, Two Tailed Test