rpa70 antibody  (Bio-Techne corporation)

Journal: Science advances

Article Title: R-loops trigger the release of cytoplasmic ssDNAs leading to chronic inflammation upon DNA damage.

doi: 10.1126/sciadv.abj5769

Figure Lengend Snippet: Fig. 6. In vitro and in vivo effect of ssDNA abrogation in Er1−/− pancreata. (A) RPA70 in nuclear and cytoplasmic extracts from wt and Er1−/− pancreata (n = 4). (B) Immunoprecipitation (IP) using anti-ssDNA in untreated or S1 nuclease–treated cytoplasmic extracts from Er1−/− pancreata for RPA70. The input (INP) and flow through (FTH) are 1/20 of the extract used (n = 3). (C) RPA70 in whole-cell extracts of wt and Er1−/− PPCs, untreated, or incubated with scrambled (sictrl) or siRNA against Rpa70 (siRPA70) (n = 4). (D) Immunostaining of ssDNA in wt and Er1−/− PPCs, untreated, or incubated with scrambled (sictrl) or siRNA against Rpa70 (siRPA70) (n = 3). (E) Immunostaining of ssDNA upon recombinant S1 nuclease transfection in untreated and MMC-treated (10 g/ml; 4 hours) wt PPCs (n = 3). (F) Immunostaining of NF-B upon recombinant S1 nuclease transfection in untreated and MMC-treated (10 g/ml; 4 hours) wt PPCs (n = 3). (G) Ifn mRNA levels in MMC-treated wt PPCs, untreated, or transfected with S1 nuclease (n = 5). The graphs in (A) and (C) represent the -TUB– or fibrillarin (FBL)–normalized expression of RPA70 levels (n.e.l.) in Er1−/− compared to corresponding wt controls, as indicated. The graphs in (D) to (F) depict the MFI per cell. All tissues and cells are derived from P15 mice. Scale bars, 5 m. Error bars indicate SEM among n ≥ 3 replicates.

Article Snippet: 53BP1 (NB100-304; IF: 1300), RPA70 (NB100-2204; WB: 1/1000), TOP1 (NBP1-30482; WB: 1/1000), and goat anti-mouse IgM 550 (NB120-9167R; IF: 1/200) were from Novus Biologicals.

Techniques: In Vitro, In Vivo, Immunoprecipitation, Incubation, Immunostaining, Recombinant, Transfection, Expressing, Derivative Assay