Fig. 6: RPPA profiling and drug screen using mouse penile cancer cells.

a Unsupervised hierarchical clustering of penile samples from mice of denoted genotypes and treatment (cisplatin) based on normalized RPPA data. One-way ANOVA identified 117 differentially expressed proteins (P < 0.05), which were used for the clustering. b Venn diagram showing the relationship of differentially expressed proteins between SA vs. WT and SAP vs. WT comparisons, with data from RPPA. two-sided Student’s t test was used to select significant proteins for each comparison (P < 0.05). c Normalized RPPA signals of selected proteins upregulated in both SA and SAP tumors compared with WT penises, with protein names and functions shown (n = 5, 8, and 4 for WT, SA and SAP, respectively). d Normalized RPPA signals of selected proteins downregulated in both SA and SAP tumors compared with WT penises (n = 5, 8, and 4 for WT, SA and SAP, respectively). e Normalized RPPA signals and ratios for phospho-Akt (S473) and total Akt in WT, SA and SAP penises (n = 5, 8 and 4 for WT, SA and SAP, respectively), validated by western blot of the tissues. f Dot plot comparing IC50 values of SA and SAP cell lines to 31 drugs, with three drugs highlighted. g, h Cell viability in vitro measured by MTT assay for indicated drugs, with IC50 noted (n = 5 for each concentration). i Western blot showing the upregulation of phospho-HER (Tyr1248 and Tyr1221) signaling in SAP tumors compared with SA tumors (n = 3 for each group). In c–e, box plots visualize the five-number summary of a data set (minimum, lower quartile, median, upper quartile and maximum). In g, h, data represent mean ± SD. ^#P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, two-sided Student’s t test.