Group differences are too small to distinguish (CCK-8 Cell Viability Assay)

Question

Accepted Answer

Treatment and control curves are nearly overlapping, error bars overlap, no statistical significance. Common causes: 1) Drug concentration was outside the effective window (too low or too high) 2) Treatment time was too short 3) Cell line is insensitive to the treatment 4) Cell state was inconsistent across wells 5) Poor experimental reproducibility Solutions: 1) Optimize drug concentration and treatment time (run a concentration gradient) 2) Test multiple time points 3) Confirm uniform cell state before treatment 4) Increase replicate count to ≥ 3 5) Cross-validate with a second readout (apoptosis, EdU, etc.)