Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: DNA Methyltransferase Gene Polymorphisms for Prediction of Radiation-Induced Skin Fibrosis after Treatment of Breast Cancer: A Multifactorial Genetic Approach

doi: 10.4143/crt.2016.256

Figure Lengend Snippet: Association analysis between polymorphisms of DNA methyltransferase genes and risk of grade ≥ 2 radiation-induced fibrosis (LENT-SOMA scale) in breast cancer patients

Article Snippet: Determination of SNPs was conducted on genomic DNA by real-time polymerase chain reaction (PCR) using the following TaqMan Pre-Designed SNP Genotyping assays (Applied Biosystems, Milan, Italy): C_27838930_10 ( DNMT1 rs2228611); C_8722920_10 ( DNMT3A rs1550117); C_7863728_10 ( DNMT3A rs7581217); C_16013055_10 ( DNMT3B rs2424908); and C_16269889_10 ( XRCC1 rs2682585).

Techniques: Significance Assay

Journal: International immunopharmacology

Article Title: The epigenetic-modified downregulation of LOXL1 protein mediates EMT in bladder epithelial cells exposed to benzo[a]pyrene and its metabolite BPDE.

doi: 10.1016/j.intimp.2024.113232

Figure Lengend Snippet: Fig. 5. DNMT3a and DNMT3b are key factors in the regulation of LOXL1 methylation modification. (A-D) The protein level of DNMT3a/3b was detected by WB assay in SV-HUC-1 cells exposed to the indicated concentrations (1 μM and 2 μM) of B[a]P/BPDE for 24 h (Fig. 5A), 1 month (Fig. 5B), 3 months (Fig. 5C), and 6 months (Fig. 5D); (E) DNMT3a knockdown construct was stably transfected into SV-HUC-1 cells, and knockdown efficiency was then determined using Western blot; (F) The efficiency of DNMT3a knockdown on downstream LOXL1 protein level was assessed using Western blot; (G) Stable transfection of DNMT3b knockdown plasmid into SV-HUC-1 cells and WB assay to detect its knockdown efficiency; (H) Western blot was used to detect the effect of DNMT3b knockdown on downstream protein LOXL1 expression.

Article Snippet: Antibodies specific against DNMT3a (sc-373905), DNMT3b (sc376043), LOXL1 (sc-166632), SLUG (sc-166476) were purchased from Santa Cruz Biotechnology (TX, USA); Antibodies specific against ECadherin (60335-1-Ig) and β-Actin (66009-1-Ig), were purchased from Proteintech (Wuhan, China).

Techniques: Methylation, Modification, Knockdown, Construct, Stable Transfection, Transfection, Western Blot, Plasmid Preparation, Expressing

Journal: International immunopharmacology

Article Title: The epigenetic-modified downregulation of LOXL1 protein mediates EMT in bladder epithelial cells exposed to benzo[a]pyrene and its metabolite BPDE.

doi: 10.1016/j.intimp.2024.113232

Figure Lengend Snippet: Fig. 6. B[a]P/BPDE exposure induces DNMT3a and DNMT3b expression in SV-HUC-1 cells by generating ROS. (A) Intracellular ROS levels were assessed using the DCFH-DA probe; (B) Quantitative analysis of fluorescence intensity; (C) Detection of intracellular ROS levels after NAC (3 mM) treatment with the ROS inhibitor using DCFH-DA probe; (D) Quantitative analysis of fluorescence intensity; (E) WB assay was performed to detect the impact of inhibitor NAC on down stream proteins expression in SV-HUC-1 cells; (F) Cell scratch assay was used to detect the impact of ROS inhibitor NAC on the migration ability of SV-HUC-1 cells.

Article Snippet: Antibodies specific against DNMT3a (sc-373905), DNMT3b (sc376043), LOXL1 (sc-166632), SLUG (sc-166476) were purchased from Santa Cruz Biotechnology (TX, USA); Antibodies specific against ECadherin (60335-1-Ig) and β-Actin (66009-1-Ig), were purchased from Proteintech (Wuhan, China).

Techniques: Expressing, Fluorescence, Wound Healing Assay, Migration

Journal: Biomolecules

Article Title: Uncovering a Novel Role of ROR1 in the Epigenetic Regulation of Tumor Suppressor Gene CREB3L1 in Triple-Negative Breast Cancer Cells.

doi: 10.3390/biom15050734

Figure Lengend Snippet: Figure 1. ROR1 knockdown decreases DNMT3A and DNMT3B expression and activity in TNBC cells. (a) Western blot analysis comparing protein levels of DNMTs in control and siROR1 in MDA- MB-231 and HCC1806 cells. (b) Quantification of Western blot analysis. In MDA-MB-231 cells, ROR1 knockdown resulted in a 0.5-fold change in DNMT3A protein levels. In HCC1806 cells, ROR1 knockdown resulted in a 0.4-fold change in DNMT3B protein levels. (c) The bar graph revealed that knocking down ROR1 expression reduces the nuclear activity of DNMT3A and DNMT3B in TNBC cells (** p < 0.01 * p < 0.05, n = 3). Statistical significance was determined through paired Student’s t-tests.

Article Snippet: Immunoprecipitation was performed overnight at 4 ◦C using DNMT3A (Cell Signaling, 57868, 1:100, Cambridge, UK) or DNMT3B (Cell Signaling, 57868T, 1:50, Cambridge, UK) antibodies.

Techniques: Knockdown, Expressing, Activity Assay, Western Blot, Control

Journal: Biomolecules

Article Title: Uncovering a Novel Role of ROR1 in the Epigenetic Regulation of Tumor Suppressor Gene CREB3L1 in Triple-Negative Breast Cancer Cells.

doi: 10.3390/biom15050734

Figure Lengend Snippet: Figure 4. CREB3L1 nuclear expression increased and nuclear DNMT3A/DNMT3B expression decreased following ROR1 knockdown. (a) Western blot analysis investigating the effect of ROR1 knockdown by siRNA on CREB3L1. (b) Quantification of Western blot analysis. ROR1 knockdown resulted in a 1.4-fold change in CREB3L1 expression in MDA-MB-231 cells and a 1.7-fold change in CREB3L1 protein expression in HCC1806 cells (* p < 0.05, n = 3). (c–f) Immunofluorescence analysis and quantification investigating how ROR1 expression affects the nuclear expression of (c,d) DNMT3A, DNMT3B, and (e,f) CREB3L1 (**** p < 0.0001, n = 30). ROR1 knockdown resulted

Article Snippet: Immunoprecipitation was performed overnight at 4 ◦C using DNMT3A (Cell Signaling, 57868, 1:100, Cambridge, UK) or DNMT3B (Cell Signaling, 57868T, 1:50, Cambridge, UK) antibodies.

Techniques: Expressing, Knockdown, Western Blot, Immunofluorescence

Journal: Biomolecules

Article Title: Uncovering a Novel Role of ROR1 in the Epigenetic Regulation of Tumor Suppressor Gene CREB3L1 in Triple-Negative Breast Cancer Cells.

doi: 10.3390/biom15050734

Figure Lengend Snippet: Figure 6. STAT3 phosphorylation had an inverse effect on CREB3L1 protein expression in TNBC. (a) Western blot analysis comparing the expression of STAT3, pSTAT3, and CREB3L1 in TNBC cells treated with scRNA to the same cell lines with ROR1 expression knocked down (siROR1). (b) Quantification of Western blot analysis (n = 3). Statistical significance was determined through paired Student’s t-test (* p < 0.05, ** p < 0.01). (c) Western blot analysis comparing the protein expression of ROR1, STAT3, pSTAT3, DNMT3A, DNMT3B, and CREB3L1 in naive MCF10A cells compared to the same cell line that has been transfected with IL-6 recombinant protein (* p < 0.05, ** p < 0.01, n = 3).

Article Snippet: Immunoprecipitation was performed overnight at 4 ◦C using DNMT3A (Cell Signaling, 57868, 1:100, Cambridge, UK) or DNMT3B (Cell Signaling, 57868T, 1:50, Cambridge, UK) antibodies.

Techniques: Phospho-proteomics, Expressing, Western Blot, Transfection, Recombinant