Journal: The Journal of Clinical Investigation
Article Title: MRE11 deacetylation by SIRT2 promotes DNA binding to facilitate DNA end resection and ATM-dependent signaling
doi: 10.1172/JCI186711
Figure Lengend Snippet: ( A ) HCT116 SIRT2 WT and KO cells were treated with 6 Gy IR, harvested after 4 hours, run on SDS-PAGE, and probed with the indicated antibodies. The image shown is representative of data from 3 separate experiments conducted independently. ( B ) Quantitation of IR-induced phosphorylation of KAP1 and CHK2 relative to total KAP1 and CHK2 protein levels. Data are represented as the mean ± SD from 3 independent experiments. ** P < 0.01 and *** P < 0.001, by 2-tail Student’s t test. ( C ) HCT116 SIRT2 KO cells expressing SFB-MRE11 WT, K393Q, and K393R were treated with or without 6 Gy IR, harvested after 4 hours, run on SDS-PAGE, and probed with the indicated antibodies. Shown is a representative image of 3 independent experiments. ( D and E ) Quantitation of phosphorylation of KAP1 and CHK2 relative to total KAP1 and CHK2 protein levels. Data are presented as the mean ± SD of 3 independent experiments. Statistical significance was determined using 1-way ANOVA followed by Dunnett’s post hoc test for multiple comparisons. **** P < 0.0001.
Article Snippet: The antibodies used in this study included SIRT2 (MilliporeSigma, 09-843; Santa Cruz Biotechnology, sc-20966), GAPDH (Santa Cruz, sc-47724), Flag (Santa Cruz Biotechnology, sc-51590), GFP (Abcam, Ab6556), pan acetyl-Lys (Santa Cruz Biotechnology, sc-8649), RPA70 (Cell Signaling Technology, 2267), P-CHK2 T68 (Cell Signaling Technology, 2661S), CHK2 (Santa Cruz Biotechnology, sc-7898), P-KAP1 (Invitrogen, Thermo Fisher Scientific, MA1-2023), and KAP1 (Bethyl Laboratories, A300-767A).
Techniques: SDS Page, Quantitation Assay, Phospho-proteomics, Expressing