FIG 5.

Endogenous ACP1 localizes to the flagellum tip. (A) mRNA levels for ACP1 and ACP2 were determined by qRT-PCR in ACP1-knockdown cells grown in the absence (−) or presence (+) of tetracycline (Tet). (B) Total protein extracts were prepared from BSF cells and from procyclic ACP1-knockdown cells (ACP1-KD) grown without (−) or with (+) tetracycline and then subjected to Western blot analysis using affinity-purified anti-ACP1 antibodies (top). (Bottom) Total protein in the same samples visualized by Coomassie staining of SDS-polyacrylamide gels. (C) Immunofluorescence of ACP1-knockdown cells grown without or with tetracycline and probed with affinity-purified anti-ACP1 antibodies. Nuclear and kinetoplast DNA are stained with DAPI (blue).