For NT and T samples, the same three reference genes were found as the most stable ones (data not shown).
We noticed that there is a large amount of FCBF lists whose sizes are rather small on CNS and Colon data compared to the other three filters, but, interestingly, this causes FCBF to be the least stable on CNS data and the most stable on Colon data.
The logistic function was chosen following earlier work that had found it the most stable for FD data, even for as few as 30 data points (Amitay, Nelson, Hawkey, Cowan, & Moore, 2006).
We used Genorm [ 29] to assess the stability of the housekeeping genes included on the Low Density array panels, and RPLPO gene expression was found to be most stable in our data.
Automated functions were employed to extract submaximal values from the smoothed data by selecting the most stable second of force data within ±5% of the target intensity, followed by extraction of the coinciding EMG amplitude.
We found 14 clusters to create the most stable divisions of the data.
GeNorm analysis (Vandesompele et al, 2002) indicated that among the reference genes, human β2-microglobulin and GAPDH were the most stable in our model (data not shown).
The L. arvalis actin-1 gene was selected as the reference gene because its expression level was most stable throughout the culture (data not shown).
The geNorm facility within Statminer identified succinate dehydrogenase subunit A as the most stable housekeeping gene, and the data were therefore normalized to succinate dehydrogenase subunit A expression.
