Fig 2: Western blot validation of specific proteins identified in the proteomic analysis.Western blot analysis was used to validate some of the targets identified on proteomic profiling. The protein expression of CDC2 was 2-fold higher in the AD cells compared to the AI tumorspheres, and PARP1, SOX2 and HDGFR were expressed > 1.5-fold higher in the AI tumorspheres compared to the AD cells. In addition, the expression levels of MIA3, RhoA, PDGFRβ, VEGFR and EGFR was also significantly (P < 0.05) higher in the AI tumorspheres, as determined with Western blot analysis. * indicates a P value < 0.05. Results represent the mean ± SEM of multiple experiments.

Fig 3: Synthetic lethality as a therapeutic paradigm in NSCLC cancer.The simultaneous mutation of RAS and upregulation of CDK1 confer complete lethality, whereas any other combination of mutations is viable.

Fig 4: CLU regulates PP2A dephosphorylation of Cdc25C and exit from mitosis Cell cycle profile of IGR‐CaP1, IGR‐CaP1‐R‐caba; and PC3, PC3‐R‐caba cells in the presence or absence of 10 nM cabazitaxel.Schema of molecular interaction between CLU and Cdc25C that regulate Cdk1/cyclin B1 activity.

Fig 5: Knockdown of CLU leads to delay in the exit from mitosis Flow cytometry for PC3 cells after transfection with siSCR or siCLU and synchronization to the M phase with thymidine/nocodazole block. DNA content was analyzed at the indicated time point after nocodazole release. Histograms show % of cells in G2/M, over 5 h period. Error bars represent mean ± SEM, n = 3, *P < 0.05, ****P < 0.0001 by ANOVA followed by Bonferroni's post hoc correction. (2 h P = 0.0142).Immunofluorescence in PC3 cells after siSCR or siCLU transfection, synchronization and nocodazole release at indicated time points. Mitotic spindle formation was assessed by confocal microscopy, using pericentrin (centrioles; red) and β‐tubulin (microtubules; green) antibodies. DNA was counterstained with (DAPI) (blue). White arrows represent segregation abnormalities. Scale bar represents 10 μm.Flow cytometry to assess phosphohistone H3 levels in the same setting as in (A). Error bar represents mean ± SEM, n = 3, ****P < 0.0001 by ANOVA followed by Bonferroni's post hoc correction.Western blot for CLU, Cdc25C, Cdk1, and cyclin B1 antibodies in PC3 cells after transfection with siSCR or siCLU, synchronization and nocodazole release at the indicated time. Vinculin was used as loading control.