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anti symmetrical dimethylarginine antibody  (Cell Signaling Technology Inc)
97
Cell Signaling Technology Inc anti symmetrical dimethylarginine antibody
A U2OS cells were transfected with a Flag‐DDX5 expression vector in the presence of siCTL or siPRMT5. Lysates were immunoprecipitated with IgG or anti‐Flag antibodies as indicated. The WCE and the bound proteins were Western‐blotted with anti‐symmetrical <t>dimethylarginine</t> antibody or anti‐Flag antibodies. B A schematic of DDX5 helicase domain and RGG/RG motifs is shown. C GST fusion of DDX5 fragments (F1‐F4) and the RK mutant used in this study. RK (*) indicates R to K substitutions of 5 arginines (R478, R482, R484, R486, and R502). D, E Coomassie Blue (left panel) and in vitro methylation assay (right panel) of indicated GST‐DDX5 fragments and the GST‐DDX5‐RK mutant. F Immunofluorescence analysis with S9.6 and anti‐Flag antibodies of U2OS cells transfected with Flag‐tagged DDX5, DDX5‐RK, or DDX5‐XD (helicase dead). Nuclear S9.6 signal was counted in both Flag‐negative and Flag‐positive cells. The Flag‐negative cells were considered as untransfected cells. The graphs shown represent the quantification with the SEM from three independent experiments. Statistical significance was assessed using one‐way ANOVA t ‐test. **** P < 0.0001. G Immunofluorescence analysis with S9.6 and anti‐Flag antibodies of U2OS cells transfected with siCTL or siDDX5‐1 and Flag‐tagged DDX5 (+WT) or DDX5‐RK (+RK) as indicated. The graphs show the average and SEM from three independent experiments. Statistical significance was assessed using one‐way ANOVA t ‐test. **** P < 0.0001. H HEK293 cells were transfected with siCTL or siDDX5. The next day, the cells that received siDDX5 were subsequently transfected with empty pcDNA3 vector (−), expression vectors encoding Flag‐DDX5 (WT), or Flag‐DDX5‐RK (RK). Forty‐eight hours later, all the cells were subjected to DRIP‐qPCR analysis. The average with the SEM from three independent experiments is shown. Statistical significance was assessed using Student's t ‐test. * P < 0.05 and ** P < 0.01; n.s.: not significant. Source data are available online for this figure.
Anti Symmetrical Dimethylarginine Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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A U2OS cells were transfected with a Flag‐DDX5 expression vector in the presence of siCTL or siPRMT5. Lysates were immunoprecipitated with IgG or anti‐Flag antibodies as indicated. The WCE and the bound proteins were Western‐blotted with anti‐symmetrical dimethylarginine antibody or anti‐Flag antibodies. B A schematic of DDX5 helicase domain and RGG/RG motifs is shown. C GST fusion of DDX5 fragments (F1‐F4) and the RK mutant used in this study. RK (*) indicates R to K substitutions of 5 arginines (R478, R482, R484, R486, and R502). D, E Coomassie Blue (left panel) and in vitro methylation assay (right panel) of indicated GST‐DDX5 fragments and the GST‐DDX5‐RK mutant. F Immunofluorescence analysis with S9.6 and anti‐Flag antibodies of U2OS cells transfected with Flag‐tagged DDX5, DDX5‐RK, or DDX5‐XD (helicase dead). Nuclear S9.6 signal was counted in both Flag‐negative and Flag‐positive cells. The Flag‐negative cells were considered as untransfected cells. The graphs shown represent the quantification with the SEM from three independent experiments. Statistical significance was assessed using one‐way ANOVA t ‐test. **** P < 0.0001. G Immunofluorescence analysis with S9.6 and anti‐Flag antibodies of U2OS cells transfected with siCTL or siDDX5‐1 and Flag‐tagged DDX5 (+WT) or DDX5‐RK (+RK) as indicated. The graphs show the average and SEM from three independent experiments. Statistical significance was assessed using one‐way ANOVA t ‐test. **** P < 0.0001. H HEK293 cells were transfected with siCTL or siDDX5. The next day, the cells that received siDDX5 were subsequently transfected with empty pcDNA3 vector (−), expression vectors encoding Flag‐DDX5 (WT), or Flag‐DDX5‐RK (RK). Forty‐eight hours later, all the cells were subjected to DRIP‐qPCR analysis. The average with the SEM from three independent experiments is shown. Statistical significance was assessed using Student's t ‐test. * P < 0.05 and ** P < 0.01; n.s.: not significant. Source data are available online for this figure.

Journal: The EMBO Journal

Article Title: Arginine methylation of the DDX 5 helicase RGG / RG motif by PRMT 5 regulates resolution of RNA:DNA hybrids

doi: 10.15252/embj.2018100986

Figure Lengend Snippet: A U2OS cells were transfected with a Flag‐DDX5 expression vector in the presence of siCTL or siPRMT5. Lysates were immunoprecipitated with IgG or anti‐Flag antibodies as indicated. The WCE and the bound proteins were Western‐blotted with anti‐symmetrical dimethylarginine antibody or anti‐Flag antibodies. B A schematic of DDX5 helicase domain and RGG/RG motifs is shown. C GST fusion of DDX5 fragments (F1‐F4) and the RK mutant used in this study. RK (*) indicates R to K substitutions of 5 arginines (R478, R482, R484, R486, and R502). D, E Coomassie Blue (left panel) and in vitro methylation assay (right panel) of indicated GST‐DDX5 fragments and the GST‐DDX5‐RK mutant. F Immunofluorescence analysis with S9.6 and anti‐Flag antibodies of U2OS cells transfected with Flag‐tagged DDX5, DDX5‐RK, or DDX5‐XD (helicase dead). Nuclear S9.6 signal was counted in both Flag‐negative and Flag‐positive cells. The Flag‐negative cells were considered as untransfected cells. The graphs shown represent the quantification with the SEM from three independent experiments. Statistical significance was assessed using one‐way ANOVA t ‐test. **** P < 0.0001. G Immunofluorescence analysis with S9.6 and anti‐Flag antibodies of U2OS cells transfected with siCTL or siDDX5‐1 and Flag‐tagged DDX5 (+WT) or DDX5‐RK (+RK) as indicated. The graphs show the average and SEM from three independent experiments. Statistical significance was assessed using one‐way ANOVA t ‐test. **** P < 0.0001. H HEK293 cells were transfected with siCTL or siDDX5. The next day, the cells that received siDDX5 were subsequently transfected with empty pcDNA3 vector (−), expression vectors encoding Flag‐DDX5 (WT), or Flag‐DDX5‐RK (RK). Forty‐eight hours later, all the cells were subjected to DRIP‐qPCR analysis. The average with the SEM from three independent experiments is shown. Statistical significance was assessed using Student's t ‐test. * P < 0.05 and ** P < 0.01; n.s.: not significant. Source data are available online for this figure.

Article Snippet: Monomethyl arginine (R*GG) (D5A12) rabbit monoclonal antibody and anti‐symmetrical dimethylarginine antibody (SDMe‐Arginine 13222S) were from Cell Signaling.

Techniques: Transfection, Expressing, Plasmid Preparation, Immunoprecipitation, Western Blot, Mutagenesis, In Vitro, Methylation, Immunofluorescence