rap1  (Cell Signaling Technology Inc)

Journal: Scientific Reports

Article Title: PKA prevents the resection of DNA double-strand breaks and favours nonhomologous end-joining

doi: 10.1038/s41598-025-23200-x

Figure Lengend Snippet: Physical interaction between 53BP1 and PKA in G2/M. ( A ) Coimmunoprecipitation in asynchronous (treated with DMSO) cells and G2/M (RO-3306)-arrested cells. Uncropped gels are shown in supplementary data. The lower panel shows the effects of DMSO (left panel) and RO-3306 (right panel) on the cell cycle distribution. ( B ) Proximity ligation assay. Upper panel: representative images of PLA with the PKAcs antibody alone (left panel), the 53BP1 antibody alone (middle panel), and both the PKAcs and 53BP1 antibodies (right panel). Scale bars: 10 µm. Lower panel: quantification (median) of at least 3 independent experiments ( p values: * < 0.05, ** < 0.01).

Article Snippet: Western blot analysis was performed to detect 53BP1and PKAcs via a rabbit anti-53BP1 antibody (1:1000, Cell Signaling, #4938) and a rabbit anti-PKAcs antibody (1:500, Cell Signaling, mAb 5842), respectively.

Techniques: Proximity Ligation Assay

Journal: Scientific Reports

Article Title: PKA prevents the resection of DNA double-strand breaks and favours nonhomologous end-joining

doi: 10.1038/s41598-025-23200-x

Figure Lengend Snippet: PLA 53P1/PKA after exposure to hydroxyurea. ( A ) Representative images of PLA with PKAcs antibody alone (left panel), 53BP1 antibody alone (middle panel), and both PKAcs and 53BP1 antibodies (right panels) at two different doses of HU. Scale bars: 10 µm. Right panels: effect of the treatment on the cell cycle distribution. ( B ) Quantification (median) of at least 3 independent experiments ( p values: * < 0.05, ** < 0.01).

Article Snippet: Western blot analysis was performed to detect 53BP1and PKAcs via a rabbit anti-53BP1 antibody (1:1000, Cell Signaling, #4938) and a rabbit anti-PKAcs antibody (1:500, Cell Signaling, mAb 5842), respectively.

Techniques:

Journal: Scientific Reports

Article Title: PKA prevents the resection of DNA double-strand breaks and favours nonhomologous end-joining

doi: 10.1038/s41598-025-23200-x

Figure Lengend Snippet: Effect of PKA on 53BP1 foci assembly after ionizing radiation (IR). ( A ) Representative image of nuclear 53BP1 foci after IR (right panel). Scale bar: 10 µm. ( B ) Kinetics of 53BP1 foci assembly after IR (2 Gy). Upper panel: silencing of PKAcs with siRNA. Lower panel: quantification of 53BP1 (means ± SEMs) foci assembly at different times after IR, with siCTRL (sicontrol) or siPKAcs (20 nM). The data represent at least 3 independent experiments ( p values: * < 0.05). ( C ) Kinetics of pCREB phosphorylation after IR. ( D ) Effect of the PKA inhibitor H89 on IR-induced pCREB. Uncropped gels are shown in supplementary data.

Article Snippet: Western blot analysis was performed to detect 53BP1and PKAcs via a rabbit anti-53BP1 antibody (1:1000, Cell Signaling, #4938) and a rabbit anti-PKAcs antibody (1:500, Cell Signaling, mAb 5842), respectively.

Techniques: Phospho-proteomics

Journal: Scientific Reports

Article Title: PKA prevents the resection of DNA double-strand breaks and favours nonhomologous end-joining

doi: 10.1038/s41598-025-23200-x

Figure Lengend Snippet: The different roles of PKA in NHEJ. At the earliest step of DSB repair, PKA promotes and/or stabilizes the loading of 53BP1 on DSBs, impairing resection and fostering NHEJ. Resection repression leads to decreased HR efficiency. However, PKA can also act at later steps of NHEJ through the nuclear translocation of DNA-PK and the expression of NHEJ factors, including ligase IV and its cofactors (XRCC4 and XLF/Cernunos).

Article Snippet: Western blot analysis was performed to detect 53BP1and PKAcs via a rabbit anti-53BP1 antibody (1:1000, Cell Signaling, #4938) and a rabbit anti-PKAcs antibody (1:500, Cell Signaling, mAb 5842), respectively.

Techniques: Translocation Assay, Expressing