Extended Data Figure 2: CaMPARI photoconversion in isolated brains.
From: Circadian clock neurons constantly monitor environmental temperature to set sleep timing
a, Temperature stimuli and LED illumination protocol for CaMPARI experiments on isolated brains. In these experiments, the temperature was increased from 23 °C to 32 °C and only two cycles of illumination were used to avoid excessive photoconversion (see Methods). b, Quantification of the ratio of red and green CaMPARI fluorescence in clock neurons from isolated brains ectopically expressing the thermosensitive channel TRPA120, which is activated by temperatures greater than 25 °C. Clock neurons ectopically expressing TRPA1 display high levels of CaMPARI photoconversion in response to heating (grey bars, number of neurons: 5 DN1ps, 4 DN2s, 5 LNds) compared to constant temperature controls (white bars, number of neurons: 5 DN1ps, 5 DN2s, 6 LNds). c, Ratio of red to green CaMPARI fluorescence in clock neurons in isolated brains in response to cooling (white bars, 63 DN1ps, 9 DN2s, 7 DN3s, 19 LNds, 20 l-LNvs) and constant temperature (grey bars, 33 DN1ps, 10 DN2s, 10 DN3s, 26 LNds, 23 l-LNvs). Histograms represent the mean ratio of red to green fluorescence ± s.e.m., three to four brains analysed for each condition. Individual data points are shown, as the sample size is less than 10. **P < 0.005, unpaired two-tailed Student’s t-test. Specific P values are reported in the Source Data for this figure.
