Index template - A Scatterplot of percentage of change in...

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A) Scatterplot of percentage of change in sleep amount during and after dTrpA1 activation from 505 Gal4 lines driving expression of dTrpA1. The solid trend line shows the inverse correlation between amount of sleep during and following dTrpA1 activation. Drivers with significantly increased sleep following dTrpA1 activation, accompanied by a lack of reduced sleep during dTrpA1 activation, are indicated in magenta and the dashed ellipse.(B) Sleep profile of wild-type (WT) > UAS-dTrpA1 flies (black circles, n = 62) versus R52B10-Gal4 > UAS-dTrpA1 (upper panel, gray squares, n = 32), R72G06-Gal4 > UAS-dTrpA1 (middle panel, cyan squares, n = 32), or R69F08-Gal4 > UAS-dTrpA1 (lower panel, magenta squares, n = 42). Sleep time was plotted in 30-min bins. White and black bars indicate 12-hr-light and -dark periods, respectively. 12-hr dTrpA1 activation at 29°C is indicated.(C and D) Sleep amount over a 12-hr period during (C) and after (D) dTrpA1 activation for WT > UAS-dTrpA1, R52B10-Gal4 > UAS-dTrpA1, R72G06-Gal4 > UAS-dTrpA1, R69F08-Gal4 > UAS-dTrpA1, and R58H05-Gal4 > UAS-dTrpA1 (n = 24) flies. Data for all lines except R58H05-Gal4 > UAS-dTrpA1 are from the same flies as in Figure 1B.(E) Heatmap of the averaged GFP signal from the expression patterns of the eight drivers indicated in magenta in Figure 1A. Image is presented using a “royal” look- up table. Scale bar, 100 μm.(F) Schematic showing organization of the ring neurons of the ellipsoid body. Upper panel depicts the location of cell bodies, lateral triangle (post-synaptic sites) and ellipsoid rings (pre- and post- synaptic sites) of the ring neurons. Lower panel illustrates ellipsoid body ring anatomy: R1, R2, R3 and R4 ring structures are indicated.(G) Whole-mount brain immunostaining of a R69F08-Gal4 > UAS-IVS-Syn21-GFP-p10 animal with anti-GFP (green) and anti-Bruchpilot (BRP, nc82, magenta). Maximal intensity of central brain is shown Scale bar, 100 μm. Bottom right inset shows a high-magnification section of the R2 ring structure. Arrowhead indicates the R2 ring. Arrow indicates the R4 ring.(H) Arousal threshold after dTrpA1 activation. The percentage of flies aroused by mild (0.1 g), moderate (0.5 g), and strong (1.2 g) mechanical rotational stimuli at ZT2 after cessation of dTrpA1 activation. Data from UAS-dTrpA1/+ (n = 96 for mild, n = 75 for moderate, n = 64 for strong), R69F08-Gal4/+ (n = 96 for mild, n = 76 for moderate, n = 48 for strong), R69F08-Gal4/UAS-dTrpA1 (n = 60 for mild, n = 89 for moderate, n = 48 for strong), R58H05-Gal4/+ (n = 96 for mild, n = 84 for moderate, n = 44 for strong) and R58H05-Gal4/UAS-dTrpA1 (n = 93 for mild, n = 48 for moderate, n = 48 for strong) flies are shown. g, relative centrifugal force.(I) Sleep propensity of UAS- dTrpA1/+, R69F08-Gal4/+, R69F08-Gal4/UAS-dTrpA1, R58H05-Gal4/+ and R58H05-Gal4/UAS-dTrpA1 flies after dTrpA1 activation. Percentage of flies falling asleep within 5 min (left panel) and sleep latency (right panel) after arousal are shown. Mean ± SEM is shown. In this and subsequent figures “ ,” “ ∗∗ ,” “ ∗∗∗ ,” and “ns” denote p < 0.05, p < 0.01, p < 0.001, and not significant $, (A) Sleep amount over a 12 hr period during (upper) and after (lower) dTrpA1
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