I have performed a single treatment (25 micro gram) of a bacterial virulence factor in presence of multiple inhibitors to see the effect in single cell line. Untreated cells, H2O2, virulence factor alone and inhibitors alone are also considered as different controls. To compare different treatments with each-other I have performed One-way ANOWA test. Is this accurate or I'm in wrong path.
Hello Sandeep Kumar
I think you can use XY instead of Column or Grouped analysis if you can work with GraphPad.
Instead of comparing means, draw dose-response curves and compare them.
Abolfazl Ghoodjani but I'm not analyzing dose response, I'm just analyzing cellular oxidative stress markers response to only 25 ug of HPSCP in presence of inhibitors for different oxidative stress markers. I hope I'm understanding your answer correctly. Abolfazl Ghoodjani
Sandeep Kumar So, it is very easy to use the same Column-tests. Pay attention to the assumptions.
It seems you made a couple of replicates (take cells, add the virulence factor, split the cells, add a different inhibitor to each well, incubate, measure -- so similar), and each series is normalized to the fluorescence reading of the control sample in this series,presumably because each time you get quite different readings between the series.
This kind of normalization not state of the art. One of its drawbacks is that you use some information twice and don't adjust the degrees of freedom accordingly and that a "bad" value in the control will spoil the entire series of measurements.
You shoud instead use a hierarchical model with a random intercept (per series). If this is not feasable you should at least use the average across all the readings within each series to normalize against.
Note: it is likely that you should analyze the log intensities, particularily when the range ob measured values is relatively large compared to the lowest (or even to the average) values.
Note: You need a correction for multiple testing. If you compare every inhibitor against the control, you should use Dunnett's procedure. If you want to do all-pairwise comparisons, you should use Tukey's HSD.
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