Fig. 7: Modulation of GWL dependency by genetic manipulation of B55α expression levels.

a Representative Western blot showing GWL and B55α protein expression levels in CHOL cancer cell lines KKK-D068, KKK-D131, KKU-055 and MMNK1. Scans from one of n = 3 independent experiments are shown. BT-549 and HCC1143 cell lines are included as exemplars of high and low cellular C-604 sensitivity, respectively. b Quantification of n = 3 independent measurements of GWL and B55α protein expression levels accompanied by predicted C-604 *ED₅₀ values. In each experiment, the recorded expression levels were scaled, with the maximum value set to 1. Points represent results of n = 3 independent experiments (circle, square, rhombus). Bars and error bars represent means and standard deviations. c The impact of C-604 on proliferation in indicated cell lines. Cell counts were determined by microscopy and normalised to the maximum values. Bars and error bars represent means and standard deviations of n = 3 independent experiments, respectively. Mean AUC values computed from normalised counts ± standard deviations are indicated. AUC – area under the curve. d Correlation scatterplot of predicted *ED₅₀ and mean AUC values (b, c) in indicated CHOL cancer cell lines (e) Representative Western blot showing GWL and B55α expression levels in WT and B55α-overexpressing U2OS cells. Scans from one of n = 3 experiments are shown. f Quantification of GWL and total B55α protein levels accompanied by predicted C-604 *ED₅₀ values in indicated cell lines. Points represent results of n = 3 independent experiments (circle, square, rhombus). Bars and error bars represent means and standard deviations, respectively. g Representative result of the colony formation assay showing WT and B55α-overexpressing (clone #12) U2OS cell lines exposed to C-604 for 72 h. Images from one of n = 3 independent experiments are shown. h Heatmap showing colony formation capacity of WT and B55α-overexpressing U2OS cells treated with C-604 for 72 h and allowed to recover for 10 days. Results of n = 3 independent experiments are shown as separate columns. Bars and error bars in the bar plot represent the means and standard deviations of C-604 ED₅₀ estimates. Points represent results of n = 3 independent experiments (circle, square, rhombus). i Correlation scatterplot of experimental C-604 ED₅₀ and predicted *ED₅₀ values in WT and B55α-overexpressing U2OS cell lines. The incorrectly predicted clones are indicated. The solid line represents the linear model based on correctly predicted sensitivities to C-604 treatment. r – Pearson correlation coefficient, R² – coefficient of determination. Predicted C-604 sensitivity was determined using the formula *ED₅₀ = 0.399 × GWL / B55α + 0.406, where GWL and B55α represent scaled GWL and B55α protein expression levels, respectively.