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argonaute 1 ago1  (Novus Biologicals)
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Novus Biologicals argonaute 1 ago1
Fig. 6. Recruitment of YY1 polycomb and RNAi machinery components. A) Bar graph of qRT-PCR for primiRNA 16, following RNA ChIP assay using YY1 antibody with or without DNase treatment in U2os pBlock-it cells. *p b 0.01 vs. input and IP:IgG; **p b 0.01 vs. input and IP:IgG. B) Bar graph of qRT-PCR for primiRNA 16, after RNA ChIP assay using YY1 or IgG and then Dicer antibodies, with or without DNase treatment in U2os pBlock-it cells. ** p b 0.001 vs. input and IP:IgG. C) qRT-PCR for primiRNA 16, after RNA ChIP assay using YY1 or IgG and then <t>Ago1</t> antibodies, with or without DNase treatment in U2os pBlock-it cells. D) qRT-PCR for primiRNA 200b/c after RNA ChIP assay with or without DNase treatment using YY1 antibody in U2os pBlock-it cells. * and **p b 0.001 vs. input and IP:IgG. E) Bar graph of qRT-PCR for primiRNA 200b/c after RNA ChIP assay with or without DNase treatment using YY1 or IgG and then Dicer antibodies in U2os pBlock-it cells.*and **p b 0.001 vs. input and IP:IgG. F) qRT-PCR for primiRNA 200b/c with or without DNasi treatment using YY1or IgG and then Ago1 antibodies in U2os pBlock-it cells. *and **p b 0.001 vs. input and IP:IgG. All data are calculated with 2−ΔΔCT method and reported as fold enrichment over non immunorecipitated RNA (input) amplified with specific set primers and represent mean values of three different experiments performed in triplicate ± S.D. RNA was also immunoprecipitated with IgG antibody and amplified with specific set primers as experimental control. G) Representative western blot analysis after immunoprecipitation of protein extracts from U2os pBlock-it cells. YY1 antibody immunopre- cipitation and western blot with Dicer antibody and vice versa. Immunoprecipitation with YY1 antibody and detection with Ago1 antibody and vice versa. IgG immunoprecipitation was used as experimental control.
Argonaute 1 Ago1, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Fig. 6. Recruitment of YY1 polycomb and RNAi machinery components. A) Bar graph of qRT-PCR for primiRNA 16, following RNA ChIP assay using YY1 antibody with or without DNase treatment in U2os pBlock-it cells. *p b 0.01 vs. input and IP:IgG; **p b 0.01 vs. input and IP:IgG. B) Bar graph of qRT-PCR for primiRNA 16, after RNA ChIP assay using YY1 or IgG and then Dicer antibodies, with or without DNase treatment in U2os pBlock-it cells. ** p b 0.001 vs. input and IP:IgG. C) qRT-PCR for primiRNA 16, after RNA ChIP assay using YY1 or IgG and then Ago1 antibodies, with or without DNase treatment in U2os pBlock-it cells. D) qRT-PCR for primiRNA 200b/c after RNA ChIP assay with or without DNase treatment using YY1 antibody in U2os pBlock-it cells. * and **p b 0.001 vs. input and IP:IgG. E) Bar graph of qRT-PCR for primiRNA 200b/c after RNA ChIP assay with or without DNase treatment using YY1 or IgG and then Dicer antibodies in U2os pBlock-it cells.*and **p b 0.001 vs. input and IP:IgG. F) qRT-PCR for primiRNA 200b/c with or without DNasi treatment using YY1or IgG and then Ago1 antibodies in U2os pBlock-it cells. *and **p b 0.001 vs. input and IP:IgG. All data are calculated with 2−ΔΔCT method and reported as fold enrichment over non immunorecipitated RNA (input) amplified with specific set primers and represent mean values of three different experiments performed in triplicate ± S.D. RNA was also immunoprecipitated with IgG antibody and amplified with specific set primers as experimental control. G) Representative western blot analysis after immunoprecipitation of protein extracts from U2os pBlock-it cells. YY1 antibody immunopre- cipitation and western blot with Dicer antibody and vice versa. Immunoprecipitation with YY1 antibody and detection with Ago1 antibody and vice versa. IgG immunoprecipitation was used as experimental control.

Journal: Biochimica et biophysica acta

Article Title: Polycomb YY1 is a critical interface between epigenetic code and miRNA machinery after exposure to hypoxia in malignancy.

doi: 10.1016/j.bbamcr.2015.01.009

Figure Lengend Snippet: Fig. 6. Recruitment of YY1 polycomb and RNAi machinery components. A) Bar graph of qRT-PCR for primiRNA 16, following RNA ChIP assay using YY1 antibody with or without DNase treatment in U2os pBlock-it cells. *p b 0.01 vs. input and IP:IgG; **p b 0.01 vs. input and IP:IgG. B) Bar graph of qRT-PCR for primiRNA 16, after RNA ChIP assay using YY1 or IgG and then Dicer antibodies, with or without DNase treatment in U2os pBlock-it cells. ** p b 0.001 vs. input and IP:IgG. C) qRT-PCR for primiRNA 16, after RNA ChIP assay using YY1 or IgG and then Ago1 antibodies, with or without DNase treatment in U2os pBlock-it cells. D) qRT-PCR for primiRNA 200b/c after RNA ChIP assay with or without DNase treatment using YY1 antibody in U2os pBlock-it cells. * and **p b 0.001 vs. input and IP:IgG. E) Bar graph of qRT-PCR for primiRNA 200b/c after RNA ChIP assay with or without DNase treatment using YY1 or IgG and then Dicer antibodies in U2os pBlock-it cells.*and **p b 0.001 vs. input and IP:IgG. F) qRT-PCR for primiRNA 200b/c with or without DNasi treatment using YY1or IgG and then Ago1 antibodies in U2os pBlock-it cells. *and **p b 0.001 vs. input and IP:IgG. All data are calculated with 2−ΔΔCT method and reported as fold enrichment over non immunorecipitated RNA (input) amplified with specific set primers and represent mean values of three different experiments performed in triplicate ± S.D. RNA was also immunoprecipitated with IgG antibody and amplified with specific set primers as experimental control. G) Representative western blot analysis after immunoprecipitation of protein extracts from U2os pBlock-it cells. YY1 antibody immunopre- cipitation and western blot with Dicer antibody and vice versa. Immunoprecipitation with YY1 antibody and detection with Ago1 antibody and vice versa. IgG immunoprecipitation was used as experimental control.

Article Snippet: IgG immunoprecipitation was used as experimental control. cross-linked RNA-protein complex was immunoprecipitated with following antibodies: YY1 (C20, Santa Cruz Biotechnology), HIF-1α (AF1935, R&D), tristetraprolin (TTP) (A-8, Santa Cruz Biotechnology), pan ubiquitine (ab7780 Abcam), trimethyl-histone H3Lys27 (07-449 Millipore), Dicer (13D6) (ab14601, Abcam) and Argonaute 1 (Ago1) (NB100-2817, Novus Biologicals) at 4 °C overnight.

Techniques: Quantitative RT-PCR, Immunoprecipitation, Control, Western Blot