The Trypanosoma brucei subpellicular microtubule array is organized into functionally discrete subdomains defined by microtubule associated proteins
Fig 5
(A) PAVE2 RNAi was induced for 2 d, after which control and PAVE2-depleted cells were fixed with paraformaldehyde and examined by epifluorescent microscopy. (B) Quantification of DNA states in control and PAVE2 RNAi cells after 1, 2, and 4 d of RNAi induction. Bars represent the mean and the error bars are the S.D. 300 cells in both control and PAVE2 RNAi conditions from N = 3 independent biological replicates were counted at each time point. One-way ANOVAs were calculated using the averages from each replicate. ****P<0.0001, ***P<0.0005, **P<0.005, *P<0.05, n.s. P>0.05. (C) Whole-mount transmission electron microscopy of control and PAVE2 RNAi cells after 1 d of PAVE2 depletion. (D) PAVE2 RNAi was induced for 1 d and cells were harvested, fixed in paraformaldehyde, and stained with anti-α-tubulin to demarcate the subpellicular array. Superplot is of the measured distance between the kinetoplast and the outer posterior edge of the array, and the posterior edge of the nucleus to the kinetoplast of 1N1K cells in control and PAVE2 RNAi conditions. 100 1N1K cells were measured for both control and PAVE2 RNAi conditions in N = 3 independent biological replicates. Unpaired two-tailed Student’s t-tests were performed using the averages from N = 3 independent biological replicates. ***P = 0.0006. n.s. P = 0.0753. (E) Control and PAVE1 RNAi cells were fixed in paraformaldehyde and stained after 1 d of PAVE1 depletion. PAVE2 does not localize to the array in the absence of PAVE1. (F) Control and PAVE2 RNAi cells were fixed in paraformaldehyde and stained after 1 d of PAVE2 depletion. PAVE1 does not localize to the array in the absence of PAVE2.