Fig. 4. (A) Growth curve of TbNRKC RNAi-induced cells. Knockdown of TbNRKC is not lethal and cell growth is similar to non-induced cultures up to 10 days after induction. (B) Isa-1 immunofluorescence signal is present in non-induced cells (-TET) on the basal body (c). 48 hours after induction, Isa-1 signal is lost in RNAi-induced cells (+TET) (d). a,b, phase-contrast images. Bar, 5 µm. (C) TbNRKC levels analysed by western blot after RNAi induction (in days). 2 days of induction (+) is sufficient to deplete cells of TbNRKC protein. No reappearance of TbNRKC protein is detected up to 10 days of induction. The transformed cell line at time zero (0) or non-induced cells (-) express similar amounts of TbNRKC protein as wild-type cells (WT). Membranes were probed with Isa-1 (upper panel) or with L8C4 as a loading control (lower panel). 2.5x10⁶ cells/well were loaded.