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Fig. 5. GFP reporter expression (schematics show constructs used) for smp-1 and smp-2 relative to ray 1 cells in wild-type and lin-1(e1275) animals. For all panels, anterior is left. (A-E,H) Ventral views; (F,G,I-L) lateral views. (A-E) Reporter genes for smp-1 and smp-2 are expressed in the male tail hook. The translational reporter gene smp-1::GFP is expressed at the cell membrane of the developing hook (A, L3 stage male (smp-1::GFP); B, DIC, arrowheads) and in adult hook cells (E, smp-1::GFP, arrowhead) in wild-type males. The smp-2p::gfp transcriptional reporter is also expressed in the hook (C, smp-2p::gfp; D, smp-2p::gfp/DIC overlay; arrowhead) in close proximity to ray 1 (D, arrows show ray 1) in wild-type adult males. No expression is detected in rays 1-6 for any reporters analyzed (smp-1p::gfp, smp-2p::gfp and smp-1::GFP). (F,G) Both smp-2p::gfp and smp-1::GFP express in rays 7, 8 and 9 (arrows). (H) Expression of smp-2p::gfp is observed in both the normal and ectopic hook (arrowheads) of lin-1 (e1275) mutant males, and in ray 7, 8 and 9 (arrows) and in the ray tail bursa (above arrows in G). (I-L) The ajm-1::GFP reporter was used to determine the position of both the ectopic hook and ray 1 in lin-1(e1275) adult males [the hook focal plane (red) and the ray focal plane (green) are shown in overlays I-L]. In developing lin-1(e1275) males, the ectopic hook (arrowhead) is located anterior to the developing ray 1 cells (I, early L3 stage; J, late L3 stage; K, L4 stage). Ray lineages are normal in lin-1(e1275) males [I, all ray cells are present (green), ray 1 cells shown by arrows]. By the late L3 stage, ray 1 cells are already found anterior to their normal position (J, arrow) and by the L4 stage, ray 1 is still further anterior (K, arrow). In adult lin-1(e1275) males, ray 1 is frequently observed anterior (arrow, class I phenotype) to its normal position relative to other rays (L, numbering), and in closer proximity to the ectopic hook position (L, arrowhead). Scale bars: 20 µm; bar in B applies to A-G, bar in K applies to H-L.

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