Effect of small TbTim knockdown on cell growth.

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Effect of small TbTim knockdown on cell growth. Effect of small TbTim knockdown on cell growth. (A) Scatterplot representation of qRT-PCR analysis of TbTim9, TbTim10, and TbTim8/13 mRNA levels in wild-type, TbTim9 RNAi, TbTim10 RNAi, and TbTim8/13 RNAi T. brucei cells after 96 h of induction with doxycycline (1.0 µg/ml). The mRNA levels of the target transcript in wild-type parasites were considered to represent 100%. Small TbTim transcript levels were normalized with the levels of the tubulin mRNA in each sample. Values were calculated from three independent replicates (****, P < 0.0001) and are represented as scatterplots with error bars for standard deviations. (B to D) Cell growth analysis of small TbTim RNAi parasites in the absence (uninduced) and presence (induced) of doxycycline (1.0 µg/ml) for 10 days. The log of the cumulative cell numbers was plotted versus time. (E) Doubling time of TbTim9 RNAi, TbTim10 RNAi, and TbTim8/13 RNAi in the absence (uninduced) and presence (induced) of doxycycline from day 2 to day 8. Data are shown as relative ratios of induced/uninduced results. The 29-13 cell line was used for comparisons. Joseph T. Smith Jr. et al. mSphere 2018; doi:10.1128/mSphere.00204-18