Fig. 6.
Association of TbARL6 with the BBSome. (A) Immunofluorescence analysis of T. brucei BSF transfected line 427/pMYCBBS1 grown in the presence of tetracycline for 24 h. Cells were incubated on ice with the lipophilic dye FM4-64 (red) to stain the flagellar pocket prior to fixing, then probed with mouse anti-myc (green) and co-stained with DAPI (blue). Bar, 5 μm. (B) Cells as in (A) above were grown in the absence (− Tet) or presence (+ Tet) of tetracycline for 72 h, then probed with mouse anti-myc (green) and rabbit anti-ARL6 (red) and co-stained with DAPI. Bar, 5 μm. (C) Total cell lysates (1 × 107 cells/lane) from BSF parental line Lister 427 (wt) and cell lines 427/pMYCBBS1 and 427/p2T7ARL6/pMYCBBS1 grown in the absence (−) or presence (+) of tetracycline for 24 h were immunoblotted and probed with mouse anti-myc and anti-NMT to monitor equal sample loading. (D) Cumulative growth of BSF transfected lines 427/pMYCBBS1 (BBS1) and 427/p2T7ARL6/pMYCBBS1 (BBS1 Arl6 RNAi) in the absence (−) and presence (+) of tetracycline, monitored over a 5 day time course (mean ± SD shown, n = 3) with statistical analysis by 1 way ANOVA (*** = p < 0.001).