Figure 3. Validation of differentially regulated proteins between CN and CR non-adherent cellular-derived patient samples.

(A) Representative Western blots of cellular proteins derived from non-adherent spheroid ascites tumor populations from CN (4 individual patient replicates) and CR (4 individual patient replicates) samples. Proteins examined are TOP2A, PYCR2 and PPL. Total protein load was obtained by staining the membrane with Deep Purple. (B) Densitometry analysis was performed using ImageStudio v5, with mean pixel intensity normalized to background. Data is expressed as mean ± SEM of n = 4 samples/group performed in triplicate. Significance was determined using Student’s t-test with *p < 0.05 considered statistically significant (C) KEGG pathway enrichment analysis of proteins identified enriched in CR. Proteins identified enriched in CR vs CN (based on label-free spectral counting, Rsc > 2, Fisher’s exact test <0.05) were functionally annotated to identify enriched biological pathways using the KEGG database. Only significantly enriched KEGG functional categories (p < 0.05) are depicted according to their protein number. The analysis yielded a strong significance for proteins associated with ‘energy metabolism’ and ‘DNA mismatch repair mechanisms’ pathways.