rat anti spi spitz rigg2a  (Developmental Studies Hybridoma Bank)

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: The following primary antibodies were then added to the membrane to incubate it: DNMT1 (Catalog #M30106M, Abmart, Shanghai, China) at 1:500 dilution, UHRF1 (Catalog #12387s, Cell Signaling Technology, Danvers, USA) at 1:1000 dilution, DNMT3A (Catalog #20954-1-AP, Proteintech, Wuhan, China) at 1:5000 dilution, DNMT3B (Catalog #26971-1- AP, Proteintech, Wuhan, China) at 1:1000 dilution, tissue inhibitor of metalloproteinase 2 (TIMP2) (Catalog #5738, Cell Signaling Technology, Danvers, USA) at 1:1000 dilution, glutathione S-transferase pi-1 (GSTP1) (Catalog #15902-1-AP, Proteintech, Wuhan, China) at 1:1000 dilution, and Ras association (RalGDS/AF-6) domain family member 1 (RASSF1) (Catalog #15902-1-AP, AbMART, Shanghai, China) at 1:500 dilution in the same buffer at 4 °C overnight.

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: The following primary antibodies were then added to the membrane to incubate it: DNMT1 (Catalog #M30106M, Abmart, Shanghai, China) at 1:500 dilution, UHRF1 (Catalog #12387s, Cell Signaling Technology, Danvers, USA) at 1:1000 dilution, DNMT3A (Catalog #20954-1-AP, Proteintech, Wuhan, China) at 1:5000 dilution, DNMT3B (Catalog #26971-1- AP, Proteintech, Wuhan, China) at 1:1000 dilution, tissue inhibitor of metalloproteinase 2 (TIMP2) (Catalog #5738, Cell Signaling Technology, Danvers, USA) at 1:1000 dilution, glutathione S-transferase pi-1 (GSTP1) (Catalog #15902-1-AP, Proteintech, Wuhan, China) at 1:1000 dilution, and Ras association (RalGDS/AF-6) domain family member 1 (RASSF1) (Catalog #15902-1-AP, AbMART, Shanghai, China) at 1:500 dilution in the same buffer at 4 °C overnight.

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

Journal: International Journal of Molecular Sciences

Article Title: Identification of a New Role of miR-199a-5p as Factor Implied in Neuronal Damage: Decreasing the Expression of Its Target X-Linked Anti-Apoptotic Protein (XIAP) After SCI

doi: 10.3390/ijms252212374

Figure Lengend Snippet: Detailed data for each antibody can be accessed at antibodyregistry.org using the provided RRID codes.

Article Snippet: anti-Placental gluthatione S-transferase (GST-pi) , , MBL International (Woburn, MA, USA) Cat# 311-H, RRID:AB_591790.

Techniques: Western Blot, Immunofluorescence, In Vitro, In Vivo