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. 2017 Dec 15;130(24):4108–4119. doi: 10.1242/jcs.207134

Fig. 5.

Fig. 5.

Complementation of CIF1-3′UTR RNAi by ectopically expressing CIF1 and its mutants. (A) Western blotting to detect the level of endogenous CIF1, which was tagged with a PTP epitope at the N-terminus, and the level of ectopically expressed CIF1 and its mutants tagged with a triple HA epitope at the C-terminus. Cells were incubated with 1.0 µg/ml tetracycline, and time-course samples were collected for western blotting. TbPSA6 level served as the loading control. (B) Immunofluorescence microscopy to detect the level of endogenous PTP-tagged CIF1 and ectopically expressed 3HA-tagged CIF1 and its mutants. Non-induced control and tetracycline-induced cells were co-immunostained with anti-Protein-A and FITC-conjugated anti-HA antibodies, and counterstained with DAPI to label nucleus and kinetoplast. (C) Immunofluorescence microscopy to monitor the localization of ectopically expressed CIF1 and its mutant in CIF1-3′UTR RNAi cells. Cells were co-immunostained with FITC-conjugated anti-HA and anti-CC2D, which labels the FAZ filament. Scale bars: 5 µm.