Figure 3. Proof-of-principle and quality control of the viability multiplexing assay.

Two cell lines stably expressing wild type (HTT-Q23) and mutant Huntingtin protein (HTT-Q79) were transiently and reversely transfected with siRNA-pools (Dharmacon) targeting the human kinome additional siRNA controls. Each siRNA-pool was transfected in two replicates, incubated for 72 h and subsequently prepared for viability multiplexing as shown in Fig. 1 A. Configuration of 384-well plates for RNAi screening. B. Image plots of scored values from CTG readout with two cell lines. C. Correlation analysis of two replicates each for both cell lines measured in all fitness indicators. Data were normalized to the plate median. Grey dots represent sample siRNAs and siRNA controls are shown in colors (positive, red; negative, blue). Coefficients of determination (R²) for all fitness indicators and cell lines are summarized in Table 1.