Fig. 2
BMP8b overexpression increased innervation of the BAT and ScW. a–d Representative images of TH immunostaining (DAB) with nuclei in blue (hematoxylin) and quantification as a percentage of image, in ScW and BAT from 5-week-old Bmp8b TG versus WT (a, b) and Bmp8b+/+ versus Bmp8b–/– (c, d) female mice; a black arrow in each image indicates a brown dot representing a parenchymal TH-positive fiber; n = 3–5. Scale bars, 20 µm (a) and 10 µm (c). e, f Representative images (e) of TH immunostaining (DAB) with nuclei in blue (hematoxylin) and quantification (f) as a percentage of image, in ScW and BAT from 5-week-old Bmp8b TG and WT female mice born and housed at 28 °C; a black arrow in each image indicates a brown dot representing a parenchymal TH-positive fiber. Scale bar, 20 µm; n = 5. g, h Representative images of primary sympathetic neurons cultured with caspase inhibitors (1 μg/ml) in the absence (control) or presence of NGF (0.1 ng/ml) and with or without BMP8 (80 ng/ml) (g). Data from a representative experiment with 50 neurons per condition: number of axon branch points per soma and total length of axon projections (h). Scale bar, 50 µm. Mean ± s.e.m. are presented; *p < 0.05, **p<0.01, ***p<0.001, compared to WT using a t-test (b, d, f)
