In general, most versions of cell cycle expression profiles are cartoons based on synchronization and bulk measurement methods, e.g.. Since the shapes of these relative expression profiles are equivalent to the outputs of state variables in mathematical models of the cell cycle, they could be used to calibrate and validate mathematical models, if they closely reflected reality – i.e., if they were based on quantitative measurements. In the best case, mathematical models should be calibrated in molecular units, and if not that, then relative units on the same scale. The relative expression of parameters determined from multi-color immunofluorescence cytometry assays, while correlated, are not quantitatively related to each other, except through a tortured path that is difficult to resolve. Here we present a method to convert multi-color data to the same relative scale. This is a step toward the goal of molecular scales. We have previously published procedures for converting data for one epitope, measured by cytometry, to molecular scales. If one of the epitopes in a multi-color assay can be converted to a molecular scale, then the procedure described herein will work to convert all of the epitopes in the assay to molecular scales. The idea here is to measure more than one epitope with indirect assays using the same secondary antibody and using cells sampled from the same experimental pot in each determination. By selecting a cell LY2835219 region in multi-variate data space in which a significant range of expression occurs for each epitope and correlating the assays through an additional measurement, the relative quantities of each epitope can be put on the same relative scale. For the work presented here, we calculated this scale for cyclins A2 and B1, using S phase as the region in which the two cyclins span ranges of expression large enough to be useful, and we used DNA content as the correlating variable. We then calculated the relative expression profiles for cyclins A2 and B1 in a multi-color assay of the same cells in which we measured cyclins A2, cyclin B1, phospho-S10- histone H3, and DNA content. The expression profiles for the cyclins were then converted to the same relative scale. The results show that the two cyclins are expressed at peak at about the same levels. The caveats here are we assume the difference between recognition of the cyclin A2 and cyclin B1 monoclonal antibodies by the secondary polyclonal antibody is negligible; we assume that the differences in affinity for each epitope by the two monoclonal antibodies are negligible, and we assume that epitope exposures are approximately the same – i.e., they are not masked in a biased manner. Given these caveats, this approach is inexact, but likely to not be far off, and in absence of other relatively good approaches, this is a first step. While the role of herbivore-induced volatiles in plant-herbivore-natural enemy interactions is welldocumented aboveground, new evidence from several systems.
The shape of the expression profile say something about the period in which a specific epitope is important act on that epitope
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