Picture from Fay DS et al. (2012) Genetics "A regulatory module controlling pharyngeal development and function in ...."

FIGURE 6. sup-37 transcriptional reporter expression. Transgenic animals carrying a his-24::mCherry reporter under the control of the sup-37 promoter show broad expression in embryos, larvae, and adults. A, C, E & G are corresponding DIC images of B, D, F & H. Initial Psup-37::his-24::mCherry expression was coincident with the onset of morphogenesis and was consistently enriched in pharyngeal cells (A-H). A premorphogenetic 200-cell-stage embryo (* in C, D) shows no expression. Scale bar in A: 10 um in A-F; 50 um in G & H.

Picture from Fay DS et al. (2012) Genetics "A regulatory module controlling pharyngeal development and function in ...."

FIGURE 7. SUP-37 translational reporter expression. A full-length isoform a SUP- 37::mCherry reporter shows expression in embryos and adult tissues. A and C are corresponding DIC images of B and D. Late-stage embryos of strain WY803 (A, B) show localization of SUP-37::mCherry in nuclei of multiple cell types. Expression in the adult pharynx of strain WY835 (C, D) show expression of SUP-37::mCherry in the multinucleate muscle groups pm3, pm4, and pm6. In contrast to the embryo, expression in the adult pharynx is both nuclear and cytoplasmic. For images C and D, anterior is to left. Scale bar in A: 10 um in A & B; 25 um in C & D.

Picture from Yi P et al. (2017) Curr Biol "Dynein-Driven Retrograde Intraflagellar Transport Is Triphasic in C.elegans ...."

(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.

Picture from Yi P et al. (2017) Curr Biol "Dynein-Driven Retrograde Intraflagellar Transport Is Triphasic in C.elegans ...."

Figure S1. Improved fluorescence imaging of C. elegans sensory cilia. Related toFigure 1 and 2.(A) Comparison of fluorescence signals in XBX-1 overexpression (OE) and knock-in (KI)lines. Fluorescence in the cell body and dendrite of the KI strain is weaker than that inthe overexpression line. Arrows indicate the cell bodies of PHA/B neurons. Arrowheadsindicate the ciliary bases of phasmid cilia. Scale bar: 5 um.(B-C) Imaging of the endogenous dynein-2 heavy chain (GFP::CHE-3) movement withan EMCCD camera and a 100x objective (red) or a cMOS camera and a 150x objective(green). A pair of phasmid cilia (numbered as 1 and 2) extend from the PHA or PHBdendritic endings and each consists of the ciliary base, middle segment (m.s.) anddistal segments (d.s.). A stack containing 200 time-lapse frames of the middlesegments were projected at maximal intensity into a single plane to show the overalllocation of multiple particles (also see Movie S1). Note with improved imageacquisition (right), two major tracks appear distinguishable, which were confirmed byplot of normalized intensity across the middle segments in (C). The distance betweenthe two tracks is ~300 nm. Scale bar: 5 um.

Picture from Yi P et al. (2017) Curr Biol "Dynein-Driven Retrograde Intraflagellar Transport Is Triphasic in C.elegans ...."

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.

Picture from Yi P et al. (2017) Curr Biol "Dynein-Driven Retrograde Intraflagellar Transport Is Triphasic in C.elegans ...."

Figure 1. Generation of Knockin C. elegans Expressing Fluorescent IFT Components(A) Schematic design of the CRISPR-Cas9-assisted knockin for C. elegans. One or two single-guide RNAs (sgRNAs) targeting the 50 or 30 ends of genes wereadded to pDD162 to generate the CRISPR-Cas9 plasmids (left), and 1 kb homology arms and fluorescent tags were inserted into pPD95.77 to generatehomology recombination (HR) templates (right). A pair of primers was designed to screen knockin progenies via PCR.(B) Summary of CRISPR targets, homology arms, and tags used for IFT gene knockin. PAM, protospacer adjacent motif.(C) Localization of the endogenously tagged IFT components in the amphid (top) and phasmid (bottom) cilia. Kymographs show particle movement along themiddle (M or m.s.) or distal segments (D or d.s.). Arrowheads indicate ciliary base and transition zone, and arrows indicate junctions between the middle and distalsegments. DHC, dynein-2 heavy chain; LIC, dynein-2 light intermediate chain. Images were taken at an exposure time of 200 ms. The scale bar represents 5 um.The bars in kymographs represent (horizontal) 5 um and (vertical) 5 s.(D) Representative images show triple-fluorescence labeling of dynein-2 (XBX-1), IFT-A (CHE-11), and IFT-B (IFT-81) complexes. The scale bar represents 5 um.(E) Summary of anterograde velocities of IFT components at the middle and distal segments. Numbers of IFT particles are shown in the brackets.(F) Representative intensity plots of IFT components along cilia. Intensities were all normalized to those at the dendrite endings.See also Figure S1.