Regulation of sleep by KIN-29 is not developmental.

Figure 1: Regulation of sleep by KIN-29 is not developmental. A) Schematic of the CRISPR/Cas9 strain construction with the DEGRON and GFP-tags inserted at the end of the kin-29 open reading frame. For panels B, D, E, G and H, the average movement quiescence in minutes ± SEM are displayed following exposure to 1500 J/m2 UVC irradiation. (+) indicates that 2 mM auxin was present during development and analysis. B) UVC–SIS is similar to wild type (WT) in auxin–exposed tir1::mRuby and kin-29::degron::GFP animals, and in tir1::mRuby; kin-29::degron::GFP (KIN-29 AID) transgenic animals in the absence of auxin. C) An image of a KIN-29 AID L4 stage animal. GFP is expressed in head neurons (white arrow) and in the intestine (white arrow head) consistent with previous reports (Lanjuin and Sengupta 2002). tir1::mRuby is expressed ubiquitously, as reported (Zhang et al. 2015). D) UVC–SIS is impaired in KIN-29 AID animals exposed to auxin from the time of hatching. E) The UVC–SIS impairment produced by continuous auxin–induced degradation of KIN-29 is similar to that of kin-29(oy38) null mutants. F) Images of adult KIN-29 AID animals never exposed to auxin (top left), exposed to auxin since hatching (top right), exposed to auxin for 4 hours as adults (bottom left), and removed from auxin for 12 hours starting at the L4 stage after exposure from hatching (bottom right). White arrows point to head neurons (green), which are visible only in the absence of auxin. Images were captured with identical camera and microscope settings. G) KIN-29 AID animals show a similar defect in UVC–SIS whether exposed to auxin at hatch or from the adult stage for four hours. H) UVC–SIS in KIN-29 AID animals on auxin from hatching through adulthood (+) or until the L4 stage, and examined 12 hours later (-12 hours). Reduction of auxin-induced degradation of KIN-29 during adulthood partially restores SIS behavior. ns = not significant, *P<0.05, **P<0.01, ***P<0.001, ****P< 0.0001, two-way ANOVA or mixed-model analysis with Tukey correction for multiple comparisons. Comparisons are made against wild-type (WT) unless otherwise indicated.