Journal: Oncotarget

Article Title: P-glycoprotein attenuates DNA repair activity in multidrug-resistant cells by acting through the Cbp-Csk-Src cascade.

doi: 10.18632/oncotarget.15065

Figure Lengend Snippet: Figure 3: Reduced DNA damage response in cisplatin-treated P-gp overexpressing cells. (A) Accumulated gH2AX in Paca-S1-P1 cells treated with cisplatin. Paca-S1-V and Paca-S1-P1 cells were treated with 50 μM cisplatin for 1 h, washed and cultured in drug-free medium for various time periods. The protein levels of γH2AX were determined by western blotting. (B) Attenuation of DNA repair protein activation by P-gp. Paca-S1-V and Paca-S1-P1 cells were treated with 50 μM cisplatin and KB and KBvin10 cells with 20 μM cisplatin for 1 h, washed, and incubated for 0, 1, 3 and 6 h. Afterward, several DDR related proteins with or without activated phosphorylation, such as ATM and pATMS1981, Chk2 and pChk2T68, Brca1 and pBrca1S1524, Nbs1 and pNbs1S343, and total proteins of Mre11, Rad50, Rad51, and FANCD2 were analyzed by western blotting. β-actin was included as a loading control.

Article Snippet: Primary antibodies against P-gp, Csk and FANCD2 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), Src, pSrcY416, pSrcY527, ATM, pATMS1981, Chk2, pChk2T68, Brca1, pBrac1S1524, Nbs1, pNbs1S343, Mre11 and Rad50 from Cell Signaling Technology (Danvers, MA, USA), Rad51 from GeneTex (Irvine, CA, USA), γH2AX from Calbiochem (San Diego, CA, USA), and Cbp and β-actin from Abcam (Cambridge, MA, USA).

Techniques: Cell Culture, Western Blot, Activation Assay, Incubation, Phospho-proteomics, Control

Journal: Nucleic Acids Research

Article Title: Marker-free quantification of repair pathway utilization at Cas9-induced double-strand breaks

doi: 10.1093/nar/gkab299

Figure Lengend Snippet: Mre11 hypomorphism alters DSB repair pathway utilization. ( A ) Schema depicting the role of Mre11-mediated end resection in the regulation of DSB repair pathway choice. ( B ) Mre11 ATLD1/ATLD1 MEFs exhibit reduced expression of the MRN complex by immunoblotting. ( C–F ) DSB repair products were identified in WT , Mre11 ATLD1 -1 and Mre11 ATLD1 -2 cells. ( C ) TMEJ-del 23 bp. ( D ) TMEJ-del 39 bp. ( E ) HR repair (HRD-500 donor) and ( F ) NHEJ-ins +1 bp. Mean ± SEM are shown. Statistical significance was assessed by two-tailed t -tests. * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001.

Article Snippet: Antibodies against Mre11 (NBP2-59677) and Rad50 (NBP2-20054) were purchased from Novus Biologicals.

Techniques: Expressing, Western Blot, Two Tailed Test