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Figure 8


Fig. 8. Overexpression of kinase-dead TbNRKC in vivo. (A, top panel) Western blot of wild-type (WT), non-induced (-) and induced cells (+) for 72 hours. The membrane was probed with Isa-1 and indicates the overexpression of mNRKC in induced cells. (A, bottom panel) Growth curve of non induced (-) and induced cells (+). (B) Basal body counts of non-induced (- TET) and induced cells (+TET). After 72 hours of induction, 48.57% of induced cells had two basal bodies (2BB) compared with 63.28% of non-induced cells. 47.04% of induced cells had four basal bodies (4BB) (two mature and two immature) compared with 35.78% of non-induced cells. In induced cells 3.63% (0% for non induced) displayed more than four basal bodies (>4BB), 0.19% (0% for non induced) displayed clustered basal bodies (Clust. BB) and 0.57% displayed mature flagella pairs (MAT. PAIRS). (C) A cell overexpressing kinase-dead TbNRKC in vivo probed with YL1/2 (b) and M25 plus DAPI (c), illustrating the four-basal-body phenotype. Bar, 5 µm.





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