Its Necessary (I have to tally the effects of these two drugs). Is it Scientifically Correct?
same Question for Transfection. Two plasmids have different vector backbones. If I have to tally the effects of these two plasmids in single assay, will it be ok just transfecting two empty vectors together in the control cell?
For the first question, I think as long as you have appropriate controls, you should be fine. you would want vehicle-free control, vehicle-1 control, vehicle-2 control, vehicle 1+2 control, Experiment 1, Experiment 2, and Experiment 1+2. Less meaningful, but potentially still valid would be: vehicle-free control, vehicle 1+2 control, experiment 1 with both equivalent concentrations of vehicles, and experiment 2 with equal concentrations of both vehicles, and experiment 1+2 with equal concentrations of both vehicles (however you can see how you may lose important comparators by doing it this way which confound you results, ex: your vehicles are synergistically toxic) Your statistics will be complicated and you will need to employ mixed effects to determine the differences between individual vehicles and their combination on cellular effects separately from experimental treatments. The other consideration, given the numbers of repeated groups you are doing, you will need to adjust your N (up) or perform post-hoc repeated measures adjustments so that you avoid false discovery. I think performing a pilot test to see the effects on vehicle(s) on cell proliferation/toxicity is warranted to get a proper sample size estimation, a priori.
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