Fig. 6: eWAT-secreted OPN circulates to the bone marrow.
From: Macrophages in epididymal adipose tissue secrete osteopontin to regulate bone homeostasis
a Representative images of immunohistochemical staining of OPN (left) and the OPN-positive stained areas (right, n = 5 biologically independent samples) of the proximal tibiae of NFD- and HFD-fed groups. Scale bar: 50 μm. b Schematic representation of the isolation of bone marrow supernatant for ELISA (left). Free OPN levels in the bone marrow supernatant of NFD- and HFD-fed mice (right, n = 6 biologically independent samples). c Relative expression of Spp1 in bone marrow over the course of feeding (n = 4 biologically independent samples). d OPN expression in bone marrow supernatant from mice with or without eWATx at week 12 of the indicated diet (n = 6 biologically independent samples). e OPN expression in the serum of the HFD-fed subgroups with and without eWATx at weeks 8, 12, and 16 (n = 5 biologically independent samples). f OPN concentration in eWAT (from NFD- and HFD-fed mice at weeks 8, 12, and 16)-derived conditioned medium (n = 5 biologically independent samples). g OPN concentration in iWAT (from NFD- and HFD-fed mice at weeks 8, 12, and 16)-derived conditioned medium (n = 4 biologically independent samples). h Ex vivo imaging (IVIS200 system) of the indicated organs and tissues collected 1 h (left part of the right box) or 24 h (right part of the right box) after unilateral eWAT injection of rOPN-FITC derived from HFD-fed mice at week 12. i, j Representative images of immunohistochemical staining of OPN (anti-human OPN antibody) in the spine, femur, tibia (proximal and distal), eWAT (ipsilaterally injected depot and contralateral depot) (i), lung, heart, liver, kidney and iWAT (j) of mice injected with rOPN-FITC into the unilateral eWAT for 1 h (top row) and 24 h (bottom row), respectively. Scale bar: 100 μm. BM: bone marrow. Images are representative of 3 independent experiments. All data are presented as mean ± SD. Two-way ANOVA with Sidak’s post-hoc test (a–c, e–g) and two-way ANOVA with Tukey’s post-hoc test (d) were used. Source data are provided as a Source Data file. See also Supplementary Table 6.
