Fig. 4

Diminished size of sympathetic chain ganglia by combination of diabetes and Hif1a deletion. A Representative images of microdissected stellate ganglia (STG) of the secondary sympathetic chain of reporter control-Ai14 and Hif1aCKO-Ai14 mice. Samples were cleared (CUBIC protocol), imaged, and reconstructed using 3D light-sheet fluorescence microscopy showing tdTomato⁺ neurons (see Additional file 2: Video S1, Additional file 3: Video S2, Additional file 4: Video S3, Additional file 5: Video S4). Scale bar 500 µm. B Representative images of the secondary sympathetic chain (the STG and four upper ganglia of the thoracic sympathetic chain) reconstructed using 3D light-sheet fluorescence microscopy (see Additional file 6: Video S5, Additional file 7: Video S6, Additional file 8: Video S7, Additional file 9: Video S8). Scale bar 1000 µm. (C) Confocal images of immunostaining for NeuN (a marker for differentiated neurons) of tdTomato⁺ reporter labeled neurons show neuronal density in transversal section of STG ganglion. Hoechst-stained cell nuclei. Scale bar 50 µm. D Quantification of the area of the STG and E the area of the sympathetic chain from STG to 4th thoracic ganglion. Data are mean ± SD (n = 3–4). F Density of NeuN⁺ cells was quantified per area of the STG (n = 4 samples per genotype, 2 section per sample). Data are mean ± SD. Statistical significance assessed by two-way ANOVA followed by post hoc Tukey ‘s multiple comparisons test **P < 0.01