Journal: Communications Chemistry

Article Title: A Hybrid compound H93 treats prostate cancer by directly binding UHRF1 and promoting protein dimerization

doi: 10.1038/s42004-025-01744-3

Figure Lengend Snippet: a Colony formation assays were used to test the inhibitory activity of H93 in human umbilical vein endothelial cells (HUVEC) or PCa cells. Experiments were performed in triplicate. (Upper) Representative images of colony formation assays are presented. (Lower) The half-maximal inhibitory concentration (IC 50 ) values of each cell line are shown as mean ± SD. b UHRF1 protein expression in HUVEC and PCa cells was assessed by immunoblotting. Experiments were performed in triplicate. c The correlation between H93 inhibitory activity and UHRF1 expression levels in PCa cells was analyzed. A scatter plot illustrates the relationship between relative UHRF1 protein levels and IC 50 values. IC 50 values were determined using a dose-response nonlinear regression model, and Pearson’s correlation coefficient ( r ) was calculated via linear regression.

Article Snippet: UHRF1 inhibitor NSC232003(Cat# HY-103236), PARP inhibitor olaparib (Cat# HY-10162) and androgen receptor inhibitor enzalutamide (Cat# HY-70002) were purchased from MCE (Shanghai, China).

Techniques: Activity Assay, Concentration Assay, Expressing, Western Blot

Journal: Communications Chemistry

Article Title: A Hybrid compound H93 treats prostate cancer by directly binding UHRF1 and promoting protein dimerization

doi: 10.1038/s42004-025-01744-3

Figure Lengend Snippet: a Co-immunoprecipitation (Co-IP) showed that H93 promotes the protein dimerization of exogenously expressed UHRF1. HEK293T cells were co-transfected with His-UHRF1 and Flag-UHRF1 plasmids, followed by treatment with 0, 5, 10, or 20 μM of H93 for 8 h. The cell lysates were incubated with protein A/G bead-bound anti-His antibody, then subjected to immunoblotting with anti-His or anti-Flag antibodies. b HEK293T cells were co-transfected with VC155-UHRF1, UHRF1-VN155 and dsRed plasmids for 48 h and then treated with H93 (0, 5, 10 or 20 μM) for 2, 4 or 6 h. The protein interaction between VC155-UHRF1 and UHRF1-VN155 was assessed by bimolecular fluorescence complementation (BiFC) assays. The length of the scale bar is 20 μm. c HEK293T cells were co-transfected with His-UHRF1 and Myc-DNMT1 plasmids, and then treated with H93 (0, 5, 10 or 20 μM) for 8 h. Cell lysates were subjected to Co-IP using an anti-His antibody, and DNMT1 protein was assessed by immunoblotting. d DU145 (Left) or VCaP (Right) cells were treated with H93 (0, 5, 10 or 20 μM) for 8 h, and the endogenous protein interaction between UHRF1 and DNMT1 was measured by Co-IP. e H93 had no significant effect on the interaction of UHRF1 and DNMT1 when the W527, R528, and R581 residues were mutated to alanine (UHRF1 3A ). HEK293T cells were co-transfected with myc-DNMT1 and His-UHRF1 WT or His-UHRF1 3A plasmids for 48 h, followed by treatment with 0 or 20 μM H93 for 8 h and subjected to Co-IP. Experiments were performed in at least triplicate, and representative images are presented.

Article Snippet: UHRF1 inhibitor NSC232003(Cat# HY-103236), PARP inhibitor olaparib (Cat# HY-10162) and androgen receptor inhibitor enzalutamide (Cat# HY-70002) were purchased from MCE (Shanghai, China).

Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Transfection, Incubation, Western Blot, Fluorescence

Journal: Communications Chemistry

Article Title: A Hybrid compound H93 treats prostate cancer by directly binding UHRF1 and promoting protein dimerization

doi: 10.1038/s42004-025-01744-3

Figure Lengend Snippet: a The binding affinity between H93 and UHRF1 SRA was measured using isothermal titration calorimetry (ITC) assays. Representative images are shown. b H93 (magenta) interacts with UHRF1 SRA (cyan) through hydrogen bonds. The final refined 2Fo-Fc electron density map for H93 is contoured at 1.0 σ and shown in gray. Key interacting residues are displayed as stick representations, with hydrogen bonds indicated by black dotted lines. c H93 is positioned within the pocket formed at the UHRF1 SRA dimerization interface. d Key interactions at the UHRF1 SRA dimer interface. UHRF1 SRA molecules (cyan and lime) and H93 (magenta) are represented as sticks, with hydrogen bonds depicted as black dotted lines.

Article Snippet: UHRF1 inhibitor NSC232003(Cat# HY-103236), PARP inhibitor olaparib (Cat# HY-10162) and androgen receptor inhibitor enzalutamide (Cat# HY-70002) were purchased from MCE (Shanghai, China).

Techniques: Binding Assay, Isothermal Titration Calorimetry

Journal: Journal of Dental Sciences

Article Title: Ubiquitin-like with phd and ring finger domains 1 as a potential therapeutic target in smoking-associated oral squamous cell carcinoma

doi: 10.1016/j.jds.2025.04.011

Figure Lengend Snippet: UHRF1 is overexpressed in oral squamous cell carcinoma (OSCC) tissues. (A–D) Gene expression analysis of UHRF1 in OSCC tissues compared to normal tissues using data from the GEO database (datasets: GSE30784 , GSE23558 , GSE74530 , and GSE37991 ). (E, F) The relative expression levels of UHRF1 in multiple OSCC cell lines were evaluated by qPCR and Western blot analysis. GAPDH was used as an internal control. All data are presented as mean ± standard deviation (SD). Statistical significance: ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001. OSCC: oral squamous cell carcinoma, UHRF1: ubiquitin-like with phd and ring finger domains 1, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: For overexpression studies, cells were transfected with a Human UHRF1 ORF cDNA expression plasmid (Cat. No. HG17896-UT, Sino Biological, Beijing, China) according to the manufacturer's protocol.

Techniques: Gene Expression, Expressing, Western Blot, Control, Standard Deviation, Ubiquitin Proteomics

Journal: Journal of Dental Sciences

Article Title: Ubiquitin-like with phd and ring finger domains 1 as a potential therapeutic target in smoking-associated oral squamous cell carcinoma

doi: 10.1016/j.jds.2025.04.011

Figure Lengend Snippet: Effects of UHRF1 knockdown on OSCC cells. (A, B) The efficiency of UHRF1 knockdown in YD38 and OEC-M1 cells was assessed by qPCR and Western blot analysis. GAPDH was used as an internal control. (C, D) The impact of UHRF1 knockdown on cell proliferation in OEC-M1 cells was evaluated using the colony formation assay. Colony numbers per well were averaged from three independent experiments. (E, F) The effects of UHRF1 knockdown on cell invasion in YD38 and OEC-M1 cells were assessed using the Transwell invasion assay. (G, H) The influence of UHRF1 knockdown on cell migration in OEC-M1 cells was analyzed using the wound healing assay. All data are presented as mean ± standard deviation (SD). Statistical significance: ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001. UHRF1: ubiquitin-like with phd and ring finger domains 1, Ctrl: untreated control; sc: scramble control; siUHRF1: UHRF1 knockdown. GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, 0 h: 0 h, 22 h: 22 h.

Article Snippet: For overexpression studies, cells were transfected with a Human UHRF1 ORF cDNA expression plasmid (Cat. No. HG17896-UT, Sino Biological, Beijing, China) according to the manufacturer's protocol.

Techniques: Knockdown, Western Blot, Control, Colony Assay, Transwell Invasion Assay, Migration, Wound Healing Assay, Standard Deviation, Ubiquitin Proteomics

Journal: Journal of Dental Sciences

Article Title: Ubiquitin-like with phd and ring finger domains 1 as a potential therapeutic target in smoking-associated oral squamous cell carcinoma

doi: 10.1016/j.jds.2025.04.011

Figure Lengend Snippet: Effects of UHRF1 overexpression on OSCC cells. (A, B) The efficiency of UHRF1 overexpression in YD10B and SCC25 cells was assessed by qPCR and Western blot analysis. GAPDH served as the internal control. (C, D) The impact of UHRF1 overexpression on cell proliferation in YD10B cells was evaluated using the colony formation assay. Colony numbers per well were averaged from three independent experiments. (E, F) The effect of UHRF1 overexpression on cell invasion in YD10B and SCC25 cells was determined by the transwell invasion assay. (G, H) Wound healing assays were performed to assess the influence of UHRF1 overexpression on cell migration. All data are presented as mean ± standard deviation (SD). Statistical significance: ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001. UHRF1: ubiquitin-like with phd and ring finger domains 1, Ctrl: control; pcmv3: empty pcmv3 vector; pcmv3-UHRF1: UHRF1 overexpression plasmid. GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, 0 h: 0 h, 24 h: 24 h.

Article Snippet: For overexpression studies, cells were transfected with a Human UHRF1 ORF cDNA expression plasmid (Cat. No. HG17896-UT, Sino Biological, Beijing, China) according to the manufacturer's protocol.

Techniques: Over Expression, Western Blot, Control, Colony Assay, Transwell Invasion Assay, Migration, Standard Deviation, Ubiquitin Proteomics, Plasmid Preparation

Journal: Journal of Dental Sciences

Article Title: Ubiquitin-like with phd and ring finger domains 1 as a potential therapeutic target in smoking-associated oral squamous cell carcinoma

doi: 10.1016/j.jds.2025.04.011

Figure Lengend Snippet: Cigarette smoke condensate increased the expression level of UHRF1 in vitro and in vivo. (A) Relative expression levels of UHRF1 in current smokers and never smokers were analyzed using the GEO database ( GSE8987 ). (B, C) qPCR and Western blot analyses showing UHRF1 expression in YD10B and SCC25 cells following treatment with 125 μM of cigarette smoke condensate for 24, 48, and 72 h. GAPDH was used as an internal control. (D) Effects of CSC treatment on body weight in nude mice. (E) Representative image of xenograft tumors illustrating tumor volume in mice treated with PBS or CSC. (F) Representative H&E staining of SCC25 xenograft tumors from both groups. Immunohistochemical (IHC) detection of UHRF1 expression in tumor tissues from the two treatment groups. Scale bars: 10 × , 100 μm; 20 × , 50 μm. All data are presented as mean ± standard deviation (SD). Statistical significance: ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001. UHRF1: ubiquitin-like with phd and ring finger domains 1, DMSO: Dimethyl sulfoxide, 24 h: 24 h, 48 h: 48 h, 72 h: 72 h, Ctrl: control, CSC: cigarette smoke condensate. GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, PBS: Phosphate buffered saline, H&E: hematoxylin and eosin.

Article Snippet: For overexpression studies, cells were transfected with a Human UHRF1 ORF cDNA expression plasmid (Cat. No. HG17896-UT, Sino Biological, Beijing, China) according to the manufacturer's protocol.

Techniques: Expressing, In Vitro, In Vivo, Western Blot, Control, Staining, Immunohistochemical staining, Standard Deviation, Ubiquitin Proteomics, Saline

Journal: Cell Communication and Signaling : CCS

Article Title: UHRF1 phase separation mediates stable inheritance of DNA methylation and promotes the proliferation of prostate cancer cells

doi: 10.1186/s12964-025-02309-6

Figure Lengend Snippet: UHRF1 expression is negatively correlated with the DNA methylation level at the promoter of silenced genes in prostate cancer (PC) cells. A Heatmap displaying gene expression of PC tissues and normal prostate tissues (right panel) and methylation levels in the promoter regions (left panel). Gene expression levels are shown as log₂-transformed transcripts per million (TPM). Methylation levels were calculated using the log 2 transformation of fragments per kilobase of transcript per million mapped reads (FPKM) within ± 1.5 kb of the transcription start site (TSS), derived from whole genome bisulfite sequencing ( GSE104789 ). B mRNA expression of frequently methylated genes ( GSTP1 , RASSF1 , and TIMP2 ) in C4-2B cells after transfection with control shRNA (sh-CTRL) or shRNA-UHRF1 (sh-UHRF1) by real-time quantitative PCR (RT-qPCR) ( n = 3). The experiment was repeated three times. C GSTP1, RASSF1, and TIMP2 protein levels in C4-2B cells transfected with sh-CTRL or sh-UHRF1 by Western blot. The experiment was repeated three times. D mRNA expression of high frequently methylated genes ( GSTP1 , RASSF1 , and TIMP2 ) in DU145 cells after transfection with sh-CTRL or sh-UHRF1 by RT-qPCR ( n = 3). The experiment was repeated three times. E GSTP1, RASSF1, and TIMP2 protein levels in DU145 cells transfected with sh-CTRL or sh-UHRF1 by Western blot. The experiment was repeated three times. F Bisulfite sequencing methylation patterns at GSTP1 and RASSF1 sequences in C4-2B and DU145 cells. CG dinucleotides are represented by circles, solid if methylated and empty if unmethylated. Percentages of methylated CG dinucleotides are shown below each pattern. G , H DNMT1 protein expression in C4-2B and DU145 cells after transfection with sh-CTRL or sh-UHRF1 by Western blot. The experiment was repeated three times. I , J DNMT1 mRNA in PC cells after transfection with sh-CTRL or sh-UHRF1 by RT-qPCR ( n = 3). The experiment was repeated three times. K UHRF1 formed nuclear condensates and was colocalized with DNMT1 in C4-2B cells. Cells were transfected with UHRF1-EGFP and DNMT1-mCherry for 48 h and imaged. mCherry and EGFP were used as controls. Data are presented as mean ± SEM by two-tailed unpaired t -test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. NS, not significant

Article Snippet: The DNMT1 and UHRF1 DNA fragments found in this study were produced by Genscript and amplified by polymerase chain reaction (PCR) using Phanta ® Max Super-Fidelity DNA Polymerase (Catalog #P505-d1, Vazyme, Nanjing, China).

Techniques: Expressing, DNA Methylation Assay, Gene Expression, Methylation, Transformation Assay, Derivative Assay, Methylation Sequencing, Transfection, Control, shRNA, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: UHRF1 phase separation mediates stable inheritance of DNA methylation and promotes the proliferation of prostate cancer cells

doi: 10.1186/s12964-025-02309-6

Figure Lengend Snippet: TSGs expression and DNMT1 localization are altered when the phase separation of UHRF1 is disrupted. A 1,6-hex disrupted droplet formation of UHRF1 that reduced the interaction between UHRF1 and DNMT1. Cells transfected with EGFP control or EGFP-tagged UHRF1 were subjected to immunoprecipitation with DNMT1 antibody. The presence of UHRF1 in the immunoprecipitate was assessed by Western blotting against an anti-GFP antibody. The experiment was repeated three times. B Enrichment of DNMT1 within ± 1.5 kb from the transcription start sites (TSSs) of all genes in the 1,6-hex treated group and control. IgG signal was used as a negative control. Genes are sorted by their CUT&Tag signal, and profiles are shown as heatmaps. C DNMT1 enrichment in all cytosine-phosphate-guanine (CpG) island regions in 1,6-hex treated (1.5% for 30 min) group and the control. IgG signal was used as a negative control. Regions were sorted by their CUT&Tag signal, and profiles are shown as heatmaps. D , E Statistical analysis of CUT&Tag data (using antibody against DNMT1, n = 3). F GSTP1 and RASSF1 mRNA expression levels in PC cells after 1,6-hex treatment (0.5% for 8 h). The experiment was repeated three times. G Bisulfite sequencing methylation patterns at the GSTP1 and RASSF1 sequences in PC cells after 1,6-hex treatment (0.5% for 8 h). The percentages of methylated CG dinucleotides are displayed beneath each pattern, with CG dinucleotides represented by circles that are solid if methylated and empty if unmethylated. Data are presented as mean ± SEM by two-tailed unpaired t -test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. NS, not significant

Article Snippet: The DNMT1 and UHRF1 DNA fragments found in this study were produced by Genscript and amplified by polymerase chain reaction (PCR) using Phanta ® Max Super-Fidelity DNA Polymerase (Catalog #P505-d1, Vazyme, Nanjing, China).

Techniques: Expressing, Transfection, Control, Immunoprecipitation, Western Blot, Negative Control, Methylation Sequencing, Methylation, Two Tailed Test

Journal: Cell communication and signaling : CCS

Article Title: UHRF1 phase separation mediates stable inheritance of DNA methylation and promotes the proliferation of prostate cancer cells.

doi: 10.1186/s12964-025-02309-6

Figure Lengend Snippet: Fig. 1 Expression and prognostic analysis of UHRF1 in prostate cancer (PC). A Differential UHRF1 mRNA expression in the tumor and normal tissues was assessed using data from GTEx and TCGA. B UHRF1 mRNA expression in paired cancer and para-cancer tissues analyzed using TCGA data. C Progression- free interval analysis of 501 PC patients based on UHRF1 expression performed using the Kaplan-Meier plotter database. D Nomogram developed to predict 1-year, 3-year, and 5-year progression-free interval in patients with PC. E Nomogram calibration for Cox regression models and fitting analysis of actual situations. F Receiver operating characteristic (ROC) curves for UHRF1 in PC and normal samples generated using data from TCGA and GTEx

Article Snippet: After that, cells were treated for the entire night at 4 °C with a 1:100 dilution of the primary antibody against UHRF1 (Catalog #21402-1-AP, Proteintech, Rosemont, USA).

Techniques: Expressing, Generated

Journal: Cell communication and signaling : CCS

Article Title: UHRF1 phase separation mediates stable inheritance of DNA methylation and promotes the proliferation of prostate cancer cells.

doi: 10.1186/s12964-025-02309-6

Figure Lengend Snippet: Fig. 2 High UHRF1 expression promotes the proliferation of prostate cancer (PC) cells. A UHRF1 protein expression in normal prostate epithelial cells (RWPE1) and PC cells (C4-2B and DU145) by Western bolt. The experiment was repeated three times. B The histogram shows the quantification of bands related to panel A (n = 3). Results are shown as mean ± SEM. C, DUHRF1 mRNA expression in PC cells in the control shRNA (sh-CTRL) and shRNA-UHRF1 (sh- UHRF1) by real-time quantitative PCR (RT-qPCR) (n = 3). The experiment was repeated three times. E, F UHRF1 protein expression in PC cells transfected with sh-CTRL or sh-UHRF1 by Western blot. The experiment was repeated three times. G, H The histogram shows the quantification of bands related to panels E and F (n = 3). Results are shown as mean ± SEM. I, J PC cell viability after transfection with sh-CTRL or sh-UHRF1 by Cell Counting Kit-8 (CCK-8) assay (n = 3). K, L Real Time Cellular Analysis (RTCA) visually showing the growth curve of wild-type PC cells (C4-2B and DU145 cells) or PC cells after transfection with sh-CTRL or sh-UHRF1 (n = 3). Results are shown as mean ± SEM. Statistics: two-tailed unpaired t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. NS, not significant

Article Snippet: After that, cells were treated for the entire night at 4 °C with a 1:100 dilution of the primary antibody against UHRF1 (Catalog #21402-1-AP, Proteintech, Rosemont, USA).

Techniques: Expressing, Western Blot, Control, shRNA, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Transfection, Cell Counting, CCK-8 Assay, Two Tailed Test

Journal: Cell communication and signaling : CCS

Article Title: UHRF1 phase separation mediates stable inheritance of DNA methylation and promotes the proliferation of prostate cancer cells.

doi: 10.1186/s12964-025-02309-6

Figure Lengend Snippet: Fig. 4 Phase separation of UHRF1 in vitro and in prostate cancer cells. A Disorder plot of UHRF1 predicted by PONDR with the VS2L algorithm. B Structure analysis of the IDR residues in the UHRF1 by MobiDB. C Representative fluorescence microscopy images of UHRF1-EGFP (10 µmol/L protein, 150 mmol/L NaCl, 10% PEG-8000). D Time-lapse micrographs of merging droplets. The concentration of UHRF1 proteins was 10 µmol/L. E Phase diagram of UHRF1 at different concentrations in the presence of different concentrations of NaCl. F Immunofluorescence assay for UHRF1 (green) in fixed C4-2B and DU145 cells. G UHRF1-EGFP formed puncta in C4-2B and DU145 cells. Cells were transfected with EGFP control or UHRF1-EGFP for 48 h and imaged. H Quanti fication of the fluorescence of UHRF1-EGFP condensates in DU145 cell after photobleaching plotted over time. Results are shown as mean ± SEM (n = 3)

Article Snippet: After that, cells were treated for the entire night at 4 °C with a 1:100 dilution of the primary antibody against UHRF1 (Catalog #21402-1-AP, Proteintech, Rosemont, USA).

Techniques: In Vitro, Fluorescence, Microscopy, Concentration Assay, Immunofluorescence, Transfection, Control

Journal: Cell communication and signaling : CCS

Article Title: UHRF1 phase separation mediates stable inheritance of DNA methylation and promotes the proliferation of prostate cancer cells.

doi: 10.1186/s12964-025-02309-6

Figure Lengend Snippet: Fig. 5 1,6-hex disrupts UHRF1 condensates and reduces UHRF1 occupancy at cytosine-phosphate-guanine (CpG) islands and promoters of PC-related genes. A Images of UHRF1-EGFP expressed in C4-2b and DU145 cells before and after treatment with 1.5% 1,6-hex for 30 min. B Enrichment of UHRF1 within ± 1.5 kb from the transcription start sites (TSSs) of all genes in the 1,6-hex treated group and control. In addition to the IgG control, the anti-UHRF1 signals in cells with UHRF1 downregulated via shRNA were also used as the negative control. Genes are sorted by their CUT&Tag signal, and profiles are shown as heatmaps. C UHRF1 enrichment in all CpG island regions in the 1,6-hex treated (1.5% for 30 min) group and the control. In addition to the IgG control, the anti-UHRF1 signals in cells with UHRF1 downregulated via shRNA were also used as the negative control. Regions were sorted by their CUT&Tag signal, and profiles are shown as heatmaps. D, E Statistical analysis of CUT&Tag data (using antibody against UHRF1). Data are presented as mean ± SEM (n = 3) by two-tailed unpaired t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. NS, not significant. F Genome browser view of UHRF1 CUT&Tag data from untreated or 1,6-hex treated (1.5% for 30 min) DU145 cells at the GSTP1 and RASSF1 locus. The Y axis shows reads per million

Article Snippet: After that, cells were treated for the entire night at 4 °C with a 1:100 dilution of the primary antibody against UHRF1 (Catalog #21402-1-AP, Proteintech, Rosemont, USA).

Techniques: Control, shRNA, Negative Control, Two Tailed Test

Journal: Cell communication and signaling : CCS

Article Title: UHRF1 phase separation mediates stable inheritance of DNA methylation and promotes the proliferation of prostate cancer cells.

doi: 10.1186/s12964-025-02309-6

Figure Lengend Snippet: Fig. 6 TSGs expression and DNMT1 localization are altered when the phase separation of UHRF1 is disrupted. A 1,6-hex disrupted droplet formation of UHRF1 that reduced the interaction between UHRF1 and DNMT1. Cells transfected with EGFP control or EGFP-tagged UHRF1 were subjected to immu noprecipitation with DNMT1 antibody. The presence of UHRF1 in the immunoprecipitate was assessed by Western blotting against an anti-GFP antibody. The experiment was repeated three times. B Enrichment of DNMT1 within ± 1.5 kb from the transcription start sites (TSSs) of all genes in the 1,6-hex treated group and control. IgG signal was used as a negative control. Genes are sorted by their CUT&Tag signal, and profiles are shown as heatmaps. C DNMT1 enrichment in all cytosine-phosphate-guanine (CpG) island regions in 1,6-hex treated (1.5% for 30 min) group and the control. IgG signal was used as a negative control. Regions were sorted by their CUT&Tag signal, and profiles are shown as heatmaps. D, E Statistical analysis of CUT&Tag data (using antibody against DNMT1, n = 3). FGSTP1 and RASSF1 mRNA expression levels in PC cells after 1,6-hex treatment (0.5% for 8 h). The experiment was repeated three times. G Bisulfite sequencing methylation patterns at the GSTP1 and RASSF1 sequences in PC cells after 1,6-hex treatment (0.5% for 8 h). The percentages of methylated CG dinucleotides are displayed beneath each pattern, with CG dinucleotides represented by circles that are solid if methylated and empty if unmethylated. Data are presented as mean ± SEM by two-tailed unpaired t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. NS, not significant

Article Snippet: After that, cells were treated for the entire night at 4 °C with a 1:100 dilution of the primary antibody against UHRF1 (Catalog #21402-1-AP, Proteintech, Rosemont, USA).

Techniques: Expressing, Transfection, Control, Western Blot, Negative Control, Methylation Sequencing, Methylation, Two Tailed Test