Fig. 2: Impact of PAKi on downstream molecular pathways and surface PD-L1 in HGSC.


a Western blot analysis of indicated proteins in Ovsaho cells treated with the indicated dose of Pan PAKi, Group II PAKi, and Group I PAKi for 4 h (left) or 48 h (right). Hsp90 and GAPDH were used as loading controls. b Heat map summarising the effects of PAKi on downstream pathways in Ovsaho cells at 4 h (upper), and 48 h (lower). Statistical significance was calculated with a two-way ANOVA; *p < 0.05. At 4 h, row factor (treatment) was significant at p = 0.0016, and the interaction between row and column factors was significant at p < 0.0001; at 48 h, row factor (treatment) and column factor (protein of interest) were both significant at p < 0.0001, and the interaction between row and column factors was significant at p = 0.0051. For each protein of interest, statistical significance was calculated with one-way ANOVA, and post hoc analyses: blank, not significant; *p < 0.05; **p < 0.01; ***p < 0.001. c Western blot analysis of indicated proteins in Kuramochi cells treated with the indicated dose of Pan PAKi, Group II PAKi, and Group I PAKi for 4 h (left) or 48 h (right). Hsp90 and GAPDH were used as loading controls. d Heat map summarising the effects of PAKi on downstream pathways in Kuramochi cells at 4 h (upper), and 48 h (lower). Statistical significance was calculated with a two-way ANOVA; *p < 0.05. At 4 h, column factor (protein of interest) was significant at p < 0.0001, and the interaction between row and column factors was significant at p = 0.0127; at 48 h, row factor (treatment) was significant at p < 0.0001, and column factor (protein of interest) was significant at p = 0.0002. For each protein of interest, statistical significance was calculated with one-way ANOVA, and post hoc analyses: blank, not significant; *p < 0.05; **p < 0.01; ***p < 0.001. e Histogram overlays show surface PD-L1 expression of Ovsaho (left) and (right) Kuramochi cells after 48 h Pan PAKi treatment. Numbered peaks represent (1) HGSC + 5 µM Pan PAKi, (2) HGSC + 5 µM Pan PAKi isotype control, (3) HGSC control (untreated cells), and (4) HGSC isotype control (untreated cells) conditions. All figures represent N = 3 biologically independent experiments.