Fig. 5: DN1as transmit temperature signals to E cells and modulate the timing of E peak.

a Examples of DN1as and E cells obtained from electron microscopy (EM) in neuPrint database. b Representative GCaMP traces (ΔF/F₀ ± SEM) of LNds in response to cooling (left, blue, n = 16) and heating (right, red, n = 16). c, f Quantification of the relative fold change of calcium activities of LNds in (b, e). Data are presented as mean in ΔF/F₀ ± SEM in the histogram; two-tailed t-test; ****p < 0.0001 (c, p = 2.03503E-28; f, left panel: p = 1.00641E-07, right panel: p = 4.11048E-14). d Representative double staining images of Anti-GFP and Anti-tdTomato in fly brains of control group (top panel) and experimental group (bottom panel). Brains were stained with anti-GFP (green) and anti-tdTomato (magenta); Scale bars, 50 μm. e representative GCaMP traces (ΔF/F₀ ± SEM) of LNds in response to cooling (left panel) and heating (right panel) in the intact fly (blue, n =  9) and DN1a silenced fly (gray, n  = 10). g, h, j Representative locomotor activity profiles of control (n = 23) and DN1a-ablated (g, n = 24) and LNd-inhibited(h, n = 24) flies at 24 °C (gray column), 18 °C (blue column) and 30 °C (j, red column, n = 24). Activity summed in 30-min bins were plotted. Bar graphs are averages ± SEM; Dark shades indicate lights off (night). i, k Quantification of E peak locomotion and E peak advance index (magenta) and E peak offset delay index (brown) in (g, j). The gray, blue and red column indicates flies’ locomotion at 22 °C or 24 °C,18 °C and 30 °C; the purple and brown plots indicate E peak advance/ offset delay index; Data are presented as mean in ΔF/F₀ ± SEM in the histogram; Statistical analysis was conducted using One-Way ANOVA followed by Tukey post-test for multiple comparisons. The letters A, B and C above the histograms denote significantly different means within each of the two groups, p < 0.05. Specific p values corresponding to this figure are reported in the Source Data. i related to (g). k related to (j).