MiCHIP data were median normalized and the log2ratios of P2 versus the corresponding senescent passage was determined.
The population doubling (PD) number of a given passage was determined by summing up the ΔPD = log2(nf/ni) for each subculture, where nf is the final number of cells in a culture and ni is the number of cells initially inoculated.
Groups of mice were inoculated with M. pulmonis, and 72 hours later, M. pulmonis numbers in lungs and nasal passages were determined.
The paracellular and transcellular passages were determined with FITC-4000 (FD-4, Sigma-Aldrich) and horseradish peroxydase (HRP Type II, Sigma-Aldrich), respectively.
The morphology of the passages is determined by the rock strata in which they are formed.
The number of cells was counted at each passage, and the number of population doublings (PDs) achieved between passages was determined by log2 (number of cells obtained/number of cells inoculated).
The number of generations traversed between passages was determined by cell counts before and after 2 days of growth on YPAD agar plates.
The concentration of S20 required for maximum virus inhibition (3 logs), while maintaining enough virus production for subsequent passages, was determined (0.5 μM S20).
The third passage cells were determined as being the most suitable for the incorporation into the alginate matrix (data not shown).
Flow cytometrically separated GFP-positive cells expressing v-Ha-Ras protein in MMH-D3, MMH-R (24-h post transfection), and MMH-R (passage 1) cells were determined by western blot analysis using the anti-v-Ha-Ras antibody.
