Fig. 5.

TAC biogenesis after recovery from p197 depletion. (A) Growth curve of γL262P 197 RNAi cells. After 5 d of p197 RNAi induction, cells were washed and grown in medium without Tet (red line). (B) Percentage of cells with kDNA and nucleus in p197 RNAi induced and uninduced trypanosomes (n ∼ 100). (C) TAC102 (green), YL1/2 (basal body, red), and DAPI (DNA, cyan) stained cells after 0 and 5 d of Tet induction (postinduction, p.i.) and 2 d postrecovery (p.r.) are shown. White arrowhead points to an example for a weak signal. (D) Quantitative analysis of TAC102 (n ≥ 106), Mab22 (n ≥ 93), or TAC40HA (n ≥ 119) in uninduced (−Tet), 5 d-induced (5 d p.i.), and 1 or 2 d after removing Tet (1 d p.r., 2 d p.r.) in γL262P p197 RNAi cells. As a control for the staining, only cells with a discernable basal body signal were used. A black bar indicates a correct localized signal; a gray bar indicates a weak signal and a light gray a mislocalized signal. (E) p197 RNAi γL262P cells were Tet induced for 3 d and stained with MitoTracker (mitochondrion, red), TAC102 (green), YL1/2 (basal body, yellow), and DAPI (DNA, cyan). (Scale bar: 2 µm.)