Figure 5. Identification of IFT-A Components IFT-139 and IFT-43 in C. elegans(A) Schematic workflow of GFP-based immunoprecipitation and sample preparation for mass spectrometry using transgenic or knockin worms.(B) Mass spectrometric results from the overexpression or knockin lines. Proteins that are homologs of subunits in kinesin-2, IFT-particle A, or IFT-particleB subcomplex were shown. ZK328.7 and C25H3.12 are putative IFT139 and IFT43 homologs, respectively.(C) Translational expression of C25H3.12::GFP in the head (arrowheads) and tail (arrows) neurons. The scale bars represent 50 um (top) and 5 um (bottom).(D) Translational expression of ZK328.7a::GFP in the head (arrowheads) and tail (arrows) neurons. The scale bars represent 50 um (left) and 5 um (right).(E) Localization of C25H3.12::GFP and ZK328.7a::GFP in the amphid (above) and phasmid (below) cilia. Kymographs show bidirectional movement along theciliary middle (M) and distal (D) segments. Arrowheads indicate ciliary base and transition zone. Arrows indicate junctions between the middle and distal segments. Images were taken at an exposure time of 200 ms. The scale bar in images represents 5 mm. The bars in kymographs represent 5 um (horizontal)
and5 s (vertical).(F) Summary of anterograde (antero.) and retrograde velocities C25H3.12 and ZK328.7a. m.s., middle segment; d.s., distal segments.See also Figure S6 and Movie S5.