FIGURE 9:
Accumulated hypophosphorylated NDK5 polypeptides in the cell body of the DN strain. (A) A representative Western blot of cell body extracts probed with anti-NDK51-201 serum. NDK5 in the DN strain P1 (lane 7) was more abundant than that in the control (lane 5) but similar to that in pf14 (x in lane 8), a control for reduced RS abundance in flagella. Negative control was ndk5 (lane 6). NDK5 bands in axonemes, by themselves or added to cell body extracts (lanes 1–4 and 9), served as markers for phosphorylated NDK5 and hypophosphorylated NDK5 that are endogenous (dot), compared with that from the transgene (triangle) or in pf14 (x). The axoneme sample loaded in lane 3 was one-sixth of the sample loaded in lane 1. (B) A Western blot probed with affinity-purified (Aff. Puri.) anti-NDK58-586 polyclonal antibody independently confirmed the hypophosphorylated state of endogenous NDK5 (dot) in the control. The band was undetectable in ndk5. The protein load was shown by Ponceau S stain (bottom).