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Fig. 8. The in vitro differentiation of CRK1+CRK2-deficient and CycE1/CYC2+CRK1+CRK2-deficient bloodstream-form cells into the procyclic form. Cells with RNAi induced for 3 days were initiated for in vitro differentiation. (A) Cell samples were collected at different time points after differentiation initiation and stained on western blot with anti-VSG221 and anti-procyclin antibodies to monitor the disappearance of VSG221 and appearance of procyclin during differentiation. Anti-{alpha}-tubulin antibody was included as a sampling control. (B) Differentiated double knockdown cells (middle panel) and triple knockdown cells (lower panel) were stained with YL1/2 antibody and compared with the procyclic-form CRK1+CRK2 knockdown cells (top panel). Note that only the cells in the top panel were stained by the antibody at the posterior ends. (C) BrdU incorporation into the differentiated double and triple knockdown cells. Note the similar outcomes to those presented in Fig. 5B and Fig. 7D on the bloodstream-form cells.





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