T. brucei growth is inhibited upon silencing of MRB8620 in glucose-poor SDM80 but not in glucose-containing SDM79. (A) Growth of PS cells in SDM79 in the presence (gray square and line) and absence (black diamond and line) of tetracycline (tet), which induces MRB8620 RNAi silencing, over a 10 d time course. The cells were diluted to the starting density of 2 × 106 cells/mL every 2 d. (B) Western blot analysis of the RNAi silencing of MRB8620-PTP by α-protein A antibody, with α-enolase shown as a loading control. Cell lysates were collected from the parental cell line (P) as well as the MRB8620 knockdowns every even day 0 to 10 d post-induction (DPI) with tetracycline. (C) Bar graph indicating the relative MRB8620 RNA levels in the RNAi-induced versus uninduced-control cells as determined by quantitative RT-PCR and normalized to 18S. Whiskers denote range of obtained relative abundancies within technical triplicates. (D–F) The same as A–C above, respectively, except that the cells were grown in SDM80.
