pchk1 (s345)  (Cell Signaling Technology Inc)

Journal: NAR Cancer

Article Title: PARP inhibitor response is enhanced in prostate cancer when XRCC1 expression is reduced

doi: 10.1093/narcan/zcaf015

Figure Lengend Snippet: Antibodies used for immunoblotting experiments

Article Snippet: pCHK1-S345 (#2348) , 1:500 , Cell Signaling Technology.

Techniques: Western Blot

Journal: The EMBO Journal

Article Title: Proteogenomic analysis reveals adaptive strategies for alleviating the consequences of aneuploidy in cancer

doi: 10.1038/s44318-025-00372-w

Figure Lengend Snippet: ( A ) Expression changes of factors involved in replication after chromosome gain (polysomic vs. WT) and after in vitro evolution (p50 vs. p0) and their comparison with the AADEPT scores (as in Fig. ). ( B , C ) Immunoblotting of the MCM2-7 subunits (Appendix Fig. shows an extended version of the immunoblotting) and the quantification in the model cell lines. The values for all subunits were normalized to the respective parental cell line and pooled together ( n : 16–18, each MCM subunit at least 2×). ( D ) Fork symmetry in the model cell lines before and after evolution and the 0.5 percentile of outliers (identified with ROUT). At least three biological replicates were analyzed, n ≥ 33 in each experiment. ( E ) Median of relative phosphorylation of the RPA2 on the S33 during S phase evaluated by flow cytometry. Three biological replicates analyzed with two technical replicates each are shown. ( F ) Quantification of relative phosphorylation of CHK1 on S345 evaluated by immunoblotting. Three biological replicates were analyzed. ( G ) Representative image and quantification of the occurrence of anaphase bridges in model cell lines. At least three biological replicates were analyzed ( n : 3–12), at least 30 anaphase cells from each were evaluated. ( H ) Representative image and quantification of the occurrence of micronuclei in model cell lines. Three biological replicates were analyzed with two technical replicates each, at least 2000 nuclei were evaluated in each experiment. All cell lines were normalized to the respective wild type. Data information: Bar plots show mean with SEM. P values were calculated using unpaired Student’s t test. Scale bars in microscopy images = 10 μm. .

Article Snippet: Rabbit anti-pChk1 (S345) WB; 1:1000 Flow cytometry; 1:1000 , Cell Signaling , 2341.

Techniques: Expressing, In Vitro, Comparison, Western Blot, Phospho-proteomics, Flow Cytometry, Microscopy

Journal: The EMBO Journal

Article Title: Proteogenomic analysis reveals adaptive strategies for alleviating the consequences of aneuploidy in cancer

doi: 10.1038/s44318-025-00372-w

Figure Lengend Snippet: ( A ) Frequency of chromosome arm gains and losses in cancer calculated from TCGA data. Black arrows indicate the frequent loss of 5q and frequent gain of 5p. ( B ) Schematic depiction of the chromosome engineering using ReDACT-TR. ( C ) Representative images of the fluorescence in situ hybridization of probes against 5p (red) and 5q (green) arms. ( D ) Chromosome-assigned shallow WGS of the parental Htr5 with Htr5p and Hdi5 cell lines. ( E ) Representative images of the clonogenic assay with quantification of the area occupied by colonies. For Htr5 and Htr5p cell lines, 16 data points were obtained from three biological replicates at 4–6 technical replicates each, for Hdi5 cell lines, 12 data points were obtained from two biological replicates at 4–6 technical experiments each. The box represents the 25th and 75th percentile with the median, whiskers show minimum and maximum values. ( F ) Mean pCHK1(S345)/CHK1 ratio determined via western Blot (at least three biological replicates; 1 to 3 technical replicates each), aphidicolin (Aph) treatment was performed for 24 h, 200 nM. Error bars represent SEM. ( G ) Survival curves of TCGA tumor patients stratified by the cooccurrence of arm-level chromosome 5 copy number gains and losses as fitted by a Cox proportional hazard model, showing the predicted survival curves of a patient of average age (58.7 years). P values are derived from Wald tests and represent the statistical significance of the hazard ratios relative to patients with copy number neutral chromosome 5. ( H ) Model of adaptation to aneuploidy in human cell lines and tumors. Data information: If not specified otherwise, P values were calculated using unpaired Student’s t test. .

Article Snippet: Rabbit anti-pChk1 (S345) WB; 1:1000 Flow cytometry; 1:1000 , Cell Signaling , 2341.

Techniques: Fluorescence, In Situ Hybridization, Clonogenic Assay, Western Blot, Derivative Assay

Journal: The EMBO Journal

Article Title: Proteogenomic analysis reveals adaptive strategies for alleviating the consequences of aneuploidy in cancer

doi: 10.1038/s44318-025-00372-w

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Rabbit anti-pChk1 (S345) WB; 1:1000 Flow cytometry; 1:1000 , Cell Signaling , 2341.

Techniques: Recombinant, Flow Cytometry, Sequencing, Control, Transfection, Software, Imaging, Cell Based Assay, Bradford Protein Assay, Western Blot, Bicinchoninic Acid Protein Assay, Peptide Fractionation