Supplementary Figure 3: LATS1 compromises CDK2 kinase activity to S3291-BRCA2 in response to stress. | Nature Cell Biology

Supplementary Figure 3: LATS1 compromises CDK2 kinase activity to S3291-BRCA2 in response to stress.

From: RASSF1A–LATS1 signalling stabilizes replication forks by restricting CDK2-mediated phosphorylation of BRCA2

Supplementary Figure 3

(a) Detection of pS3291-BRCA2 in lysates of HT1080 cells treated with the indicated siRNAs and expressing Myc-LATS1 or control plasmid. Line graphs represent time course measurement of pS3291-BRCA2 levels at indicated times post 4 Gy γIR in cycling cells (bottom) or nocodozol treated cells (top). In pS3291-BRCA2 quantification plots, error bars represent the variation in the densitometry of the representative image. Quantification of cycling cells also presented in Fig. 2e in a different scale. (b) In vitro kinase assay of immunoprecipitated CDK2 from Lats1+/+ and Lats1−/− MEFs incubated with C-terminal BRCA2 purified peptide (His-TR2) in the presence of 32[P]ATP. Reactions were resolved by SDS polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene fluoride (PVDF) membrane. The membrane was visualized by phosphoimage detection (PMI imager, Bio-Rad) and probed with the indicated antibodies.

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