Fig. 6: Naturalistic moon- and sunlight impact differently on L-Cry localization and levels.

a,a′,a″ Overview of sampling timepoints. 16h day (light) and 8h night (dark or moonlight) per 24 h, with 8 nights of moonlight per month. NM and FM-1 have the same light regime, but are named differently for accuracy as they refer to different days relative to the lunar month. b relative L-Cry levels at indicated timepoints, as determined by western Blot. Individual data points as well as mean ± SEM are shown. Ordinary one-way ANOVA: p < 0.0001; adjusted p-values of Tukey’s multiple comparison test: FM-1 zt0–10 min vs FM-1 zt 8: p < 0.0001, FM7 zt0–10 min vs FM7 zt8: p < 0.0001, FM-1 zt0–10 min vs FM7 zt 0–10 min: p = 0.9141. c Representative Western Blot used for quantification in (b), see Supplementary Fig. 7 for all other. In addition, in the context of further investigations such Western blots of identical or similar conditions were performed completely independently meanwhile more than three times with highly consistent results. d P. dumerilii head. Dashed ovals designate the oval-shaped posterior domains between the posterior eyes. Green dots: L-Cry+ cells. ae, anterior eye; pe, posterior eye. e–g Confocal single layer (1.28 µm) images of worm heads stained with anti-L-Cry antibody (green) and HOECHST (magenta: nuclei). White rectangles: areas of the zoom-ins presented below. e′–g‴ zoomed pictures of the areas depicted in e–g. Arrows: predominant nuclear L-Cry, arrowheads: predominant cytoplasmic L-Cry. Scale bars: 10 µm. Overview images with nuclear stain: Supplementary Fig. 8a–c. For a complete set of examples from a randomly chosen experimental repetition see Supplementary Fig. 10. In addition, in the context of further investigations such immunohistochemical stainings of worm heads from similar conditions were performed completely independently meanwhile more than three times with highly consistent results. h quantification of subcellular localization of L-Cry as nuclear/cytoplasmic ratio at indicated timepoints. Individual data points as well as mean ± SEM are shown. p values: two-tailed t-test. NM zt0 –10 min vs NM zt0 + 10 min: p = 0.0019, NM zt0 –10 min vs FM7 zt0–10 min: p = 0.3837, NM zt0 + 10 min vs FM7 zt0 −10 min: p < 0.0001. Biological replicates: NM zt0 −10 min n = 22; NM zt0 + 10 min n = 18; FM7 zt0 −10 min n = 16. For quantification as categorical data, see Supplementary Fig. 8a′–f.