Regulators of Trypanosoma brucei Cell Cycle Progression and Differentiation Identified Using a Kinome-Wide RNAi Screen
Figure 5
RDK2 is a repressor of BSF to PCF differentiation.
A: Predicted domain structure of RDK2. PH: pleckstrin homology domain. B: Growth curves showing cumulative cell counts over time following tetracycline (Tet) induction (+) or not (−) of RDK2 RNAi in culture. Cell densities were maintained between 105 and 106 cells ml−1. Error bars indicate the standard deviations around the means of three technical replicates. Inset: qRT-PCR analysis of RDK1 mRNA knockdown following RNAi induction. C: quantification of EP procyclin-expressing cells by flow cytometry following induction of RDK2 RNAi. RNAi of RDK2 was induced for 24 and 48 hr by addition of tet and the percentage of EP-procyclin positive cells detected by flow cytometry after treatment with 250 µM 8-pCPT-cAMP (D), cold shock treatment of cells at 27°C (E) or incubation with 150 mM BZ3 (F) for the final 24 hr of each RNAi induction.