Four hours before the end of each time point, 20 μL of 12 mM MTT was added to each well.
GO list of each time point mode is loaded separately and collected graphs are then edited by normal image processing program.
An aliquot of each time point (15, 30, 60, and 90 min, 100 μL) was injected into the analytical radio-HPLC system for analytics.
In live imaging assays, computer-assisted cell tracking showed locomotion characteristics of SMCs; reflection and fluorescent confocal microscopy and spatial reconstruction images of each time point showed detailed morphologic changes of collagen fibers and spatial collagen SMC interactions.
Four replicates of each time point were completed.
At the collection of each time point, cells were washed twice with PBS.
Error bars represent the standard deviation calculated from the triplicate analysis of each time point.
Relative band intensity of each time point was measured and set in relation to either PKD2 or actin loading control as indicated in the figure legend.
For biological replicates of each time point, nearly all of probe sets were fallen along the diagonal of plots, indicating no major variation.
At the end of each time point, cells were washed twice with PBS and with DMEM supplemented with 3% stripped FCS with or without 17βE2 for variable periods of time.
The samples of each time point (Day 1, 7, 14, 21, 28 and 35) were rejuvenated and used for analysis of 2,3-DPG levels, pH, free Hb levels, free K+ and Na+ concentrations, MCV and MCH.
