This clustering indicated that the cell line, and then inhibitor, and finally the ligand treatment imparted the most substantial changes in the cells in the y dimension. In the dimension, the data indicated that the change by time point tended to cause the most substantial re sponse in phosphoprotein levels. For Navitoclax manufacturer each treatment, biological duplicates were measured and the absolute percentage difference between the two replicates was determined. A mean difference of 20. 4% was observed across all cell lines which when compared to the finding that the phospho sites varied by approximately 670% on average over un treated controls, was considered an acceptable amount of error.

Regression analysis correlating phosphoprotein measurements to cell survival in androgen depleted conditions In an attempt to understand how the alterations in signal ing may lead to variations in survival outcomes in cells grown in androgen Inhibitors,Modulators,Libraries depleted conditions, we built a statis tical model using PLS regression. The Inhibitors,Modulators,Libraries data was arranged so that the phosphoprotein data was regressed against the survival data using PLS regression on the complete data set of 8 phosphoproteins, at 3 time points, using 3 cell lines, with 6 treatments. After calculating the model parameters the leave one out cross validated R2 value was determined to be 0. 616 with 3 latent variables, and the predicted versus measured survival values were plotted. Additional latent variables beyond three had marginal explanatory power due to the fact that the majority of the variation in the X matrix could be described in terms of these latent variables, therefore three components were used for all analyses.

When this calculated R2 value was compared to the mean R2 value calculated from randomized models we observed that this model was 6. 36 standard deviations above the mean randomized value of 0. 1847 corresponding Inhibitors,Modulators,Libraries to a P value less Inhibitors,Modulators,Libraries than 0. 0001. Inhibitors,Modulators,Libraries This result indicates that this model can correlate to survival significantly better than by random chance. Upon determining that this model was significantly more accurate than a randomized model, we examined the regression coefficients to determine weights calculated on the different phosphoproteins. Consistently positive coefficients for p Erk were noted, as well as consistently increased p RPS6 across all time points. p JNK regression coefficients were negative at all time points along with p Akt and p Stat3.

p GSK3 additionally had minimal early Tipifarnib order and late time point regression coefficients, how ever had a substantially increased 4 hour regression coefficient. In order to better assess the contribution of the regres sion coefficients to the model outcome the absolute value of the coefficients was taken for each time point and the mean plotted for each phosphoprotein in descending order. From this, p Erk was determined to most strongly contribute to the model, followed by p RPS6 and p JNK.