Additional file 5: Figure S4.
Two compartment modelling of CXCR4 trafficking. (A) Real time PCR for SDF-1: Expression of SDF-1 in fMSC (n = 4) shows SDF-1 and CXCR4 transcripts relative to the housekeeping gene GAPDH. B) Kinetics of CXCR4 exocytosis in fMSC after treatment with endocytosis inhibitors: Cells were treated with 80 μM blebbistatin or 80 μM dynasore then fixed and stained with anti-CXCR4. Surface expression was determined by flow cytometry and data fitted to a two-compartment model of endocytosis (same data as Figure 1C). C) Fit of CXCR4 surface expression data for naïve (kr = 0.04 min−1, ke = 0.79 min−1), blebbistatin- (kr = 0.04 min−1, ke = 0.078 min−1, r2 = 0.89) and dynasore-treated cells (kr = 0.03 min−1, ke = 0.09 min−1, r2 = 0.91) to the two compartment model. Simulation was initiated with cytoplasm (endosomes) containing the entire cellular CXCR4. Response of surface expression to changes in the endocytosis rate (ke): kr was maintained at 0.04 min−1 while ke was varied from 0.79 min−1 to 0.02 min−1. D) Data that was used in the mathematical modelling of CXCR4 trafficking.
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Pelekanos et al. BMC Cell Biology 2014 15:15 doi:10.1186/1471-2121-15-15