4.11. Calculation of IC50 / EC50 / CC50 results and statistical analysis.

To more easily distinguish inhibition results between the various assays, we represent the inhibitor concentrations at half-maximal effects as “IC₅₀” for protein-based biochemical assays, “EC₅₀” for cancer cell viability, migration and clonogenicity assays, and “CC₅₀” for non-cancerous cell cytotoxicity assays. All I/E/CC₅₀ results reported are averages of values determined from individual dose-response curves in replicate assays as follows: 1) Individual I/E/CC₅₀ values from replicate assays were first log-transformed and the average log(I/E/CC₅₀) values and standard deviations (SD) calculated; 2) Replicate log(I/E/CC₅₀) values were evaluated for outliers using the ROUT method in GraphPad Prism (Q of 10%); and 3) Average I/E/CC₅₀ values were then back-calculated from the average log(I/E/CC₅₀) values. For compounds where log(I/E/CC₅₀) values were greater than the maximum compound concentrations tested (i.e. >1.8, >2.0, and >2.4 – or >63, >100, and >250 μM respectively), results were represented as 0.1 log units higher than the maximum concentrations tested (i.e. 1.9, 2.1 and 2.5 – or 79, 126, and 315 μM, respectively), so as not to overly bias comparisons because of the unavailability of definitive values for these inactive compounds. Correlation factors were calculated via Pearson or Spearman analyses of log-transformed values using GraphPad prism. Significance was determined with p value as follow p > 0.05 (ns), p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***) and p ≤ 0.0001 (****).