Figure 2.
FL and ML kinesin-2 polypeptides expressed in mouse photoreceptors. A and B) Schematic representation of mouse KIF3A (A) and KIF17 (B) domain structures. Motor (blue), coiled-coil (CC; red), and tail (yellow) domains are indicated. KIF3A and KIF17 FL and ML constructs carry N-terminal fluorescent (mC or eGFP) and myc tags. C and D) Expressions of mC- and myc-tagged FL KIF3A [mC-KIF3A (C)] and ML KIF3A [mC-KIF3A-ML (D)]. Note that in 1 mo AAV-transduced retina, mC-KIF3A was absent in OS (C, inset). E) mC-KIF17 is expressed in photoreceptor IS and ONL; note that mC-KIF17 particles line up along OS axoneme (E, inset, arrows). F and G) Motorless KIF17 mutant mC-KIF17-ML1 localizes at IS and ONL (F); mC-KIF17-ML2 accumulates almost exclusively within ONL nuclei (G). Nuclear envelopes are defined by lamin B1 labeling (F, G, insets, green). H) Quantitation of photoreceptor OS lengths at 1 mo as index of degeneration: mC-KIF3A (lane 1), mC-KIF3A-ML (lane 2), mC-KIF17 (lane 3), mC-KIF17-ML1 (lane 4) and mC-KIF17-ML2 (lane 5) (n = 5, P < 0.01).