Roportional to the number of DSBs and that their appearance and disappearance correlate quite nicely together with the kinetics of DSB repair (Balajee and Geard, 2004; Olive, 2011). Because antibodies against -H2AX in C. elegans are certainly not offered, we couldn’t examine the formation of -H2AX foci in N2 or brc-1 mutants. In summary, we’ve got shown that the dissected mitotic germline region could be applied for the comet assay in C. elegans and that DNA strand breaks might be detected by glyoxal or neutral comet assays. These assays is often utilized for the detection of DNA strand breaks induced by other DNA damaging agents and for measuring DNA repair kinetics in C. elegans. Note: Supplementary information is readily available around the Molecules and Cells site (molcells.org).ACKNOWLEDGMENTSThis perform was supported by the grant of University of Ulsan (2015) to B. Ahn.Chronic myeloid leukemia (CML) is often a myeloproliferative disorder triggered by the constitutively active tyrosine kinase BCRABL, the outcome in the reciprocal chromosomal translocation (9:22) (q34;q11) (Deininger et al., 2000; Pane et al., 1996). BCR-ABL abnormally activates a number of signal pathways, inAffiliated Hospital of Integrated Standard Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing 210028, China, two Salmonella Inhibitors targets Laboratory of Cellular and Molecular Biology, Jiangsu Province Academy of Classic Chinese Medicine, Nanjing 210028, Jiangsu, China, 3 Key Laboratory of Genomics and Precision Medicine, China Gastrointestinal Cancer Study Center, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 100101, China Correspondence: [email protected] (CC); [email protected] (Computer) Received 25 January, 2016; revised 23 June, 2016; accepted 10 November, 2016; published on-line 13 December, 2016 Keywords and phrases: BCR-ABL, cantharidin, chronic myeloid leukemia, imatinib resistanceMATERIALS AND METHODSReagents and antibodies CTD (98 or greater purity) was bought from Shanghai Shifeng Biological Technology Enterprise (China). CTD was dissolved in DMSO as a 40 mM stock answer and stored at 20 . Primary antibodies against pCdc2, Cdc2, Cdc25c, cyclin B1, cyclin D1, pSTAT5, STAT5, pAKT, AKT, pERK1/2,eISSN: 0219-1032 The Korean Society for Molecular and Cellular Biology. All rights reserved. That is an open-access write-up distributed below the terms of your Creative Commons Attribution-NonCommercial-ShareAlike three.0 Unported License. To view a copy of this license, take a look at http://creativecommons.org/licenses/by-nc-sa/3.0/.Cantharidin Overcomes Imatinib Resistance in CML Xiaoyan Sun et al.ERK1/2, PARP1, cPARP, and pH3 (Ser10) have been purchased from Cell Signal Technologies, Inc. (USA). Antibodies against GAPDH and BCR were purchased from Santa Cruz Biotechnology (USA). Antibody against H2AX was from EMD Millipore (USA). Imatinib and CGK733 had been bought from Selleck (USA). Caffeine was purchased from Amquar Biology (AMQUAR Bio., USA). RPMI 1640 medium, fetal bovine serum (FBS), penicillin G and XY028-133 Data Sheet streptomycin had been offered by Gibco (USA). All of the other reagents used in this study have been of analytical grade. Cells and cell culture Human CML cell line K562 was purchased from Cell Bank of Shanghai Institute of Biochemistry and Cell Biology. K562 cells had been cultured in RPMI 1640 medium supplemented with 10 FBS. The imatinib-resistant cell line, K562R, was kindly offered by Guangbiao Zhou (Institute of Zoology, Chinese Academy of Sciences, China). K562R cells had been cultured in RPMI 1640 medium supplemented with ten FBS, a.
