Extended Data Fig. 2: Acute heat exposure induces cFOS expression in tanycytes.

a. Left: Confocal micrographs showing cFOS immunoreactivity (in cyan) at −1.94 mm (relative to bregma) after 1h exposure to either 40 °C or 4 °C vs. 25 °C, used as control, in male (♂) mice. The dorsomedial hypothalamic nucleus (DMH), the ventromedial hypothalamic nucleus (VMH) and the arcuate nucleus (ARC) were indicated. Arrowheads point to cFOS⁺/vimentin⁺ (the latter marker in magenta) tanycytes. Scale bar = 70 µm. Right: Quantitative analysis of cFOS⁺ α-tanycytes. Data in bar graphs represent means ± s.e.m. with individual data points shown as circles; n = 3/condition; one-way ANOVA: F = 16.34; p = 0.004. Bonferroni’s multiple comparison: 25 °C vs. 40 °C: t = 5.102; p = 0.007; 25 °C vs. 4 °C: t = 0.3189; p ~ 1.0 (n.s., non-significant); 40 °C vs. 4 °C: t = 4.783; p = 0.009. b. Left: Confocal micrographs showing cFOS immunoreactivity (in cyan) at −2.30 mm (relative to bregma) after 1h exposure to 40 °C or 4 °C vs. 25 °C, used as control, in male (♂) mice. Arrowheads point to cFOS⁺/vimentin⁺ (the latter marker in magenta) α-tanycytes. Scale bar = 70 µm. Right: Quantitative analysis of cFOS⁺ α-tanycytes. Data in bar graphs show means ± s.e.m. with individual data points depicted as circles; n = 3/condition; one-way ANOVA: F = 30.460; p < 0.001. Bonferroni’s multiple comparison: 25 °C vs. 40 °C: t = 7.590; p < 0.001; 25 °C vs. 4 °C: t = 2.219; p = 0.205; 40 °C vs. 4 °C: t = 5.372; p = 0.005. c. Right: Confocal micrographs of cFOS⁺ α-tanycytes (in cyan) at −1.94 mm (relative to bregma) after 1h exposure to 40 °C vs. 25 °C in female (♀) mice. Arrowheads denote cFOS⁺/vimentin⁺ tanycytes (in magenta). Scale bar = 100 µm. Right: cFOS⁺ α-tanycytes at −1.94 mm (relative to bregma) at the temperatures indicated. Data in bar graphs represent means ± s.e.m. with individual data points shown as circles; n = 4/condition; t = 14.700; p < 0.0001; Student’s t-test. d. Left: Confocal micrographs with cFOS immunoreactivity (in cyan) at −2.30 mm (relative to bregma) following the temperature manipulations as indicated in females (♀). Arrowheads point to cFOS⁺/vimentin⁺ α-tanycytes (in magenta). Scale bar = 50 µm. Right: cFOS⁺ α-tanycytes at −2.30 mm. Data in bar graphs are means ± s.e.m. with individual data points shown as circles; n = 4/condition; t = 6.331; p < 0.001; Student’s t-test.

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