Figure 5.

Analysis of ITA7 levels and spatial distribution in KO and WT adipose tissue. Immunohistochemical confocal images of fixed adipose tissue from (A) KO mice and (B) WT mice. Formaldehyde-fixed tissues were stained with antibodies against ITA7. Endothelial cells were counterstained with isolectin (red), the fatty acid with BODIPY (green) and the nucleus with DAPI (blue). (C) ITA7 volume was 64% lower in KO tissue compared to WT (1338 ± 296 μm³ in KO vs. 37,013 ± 409 μm³ in WT, p < 0.0001). (D) There is not significant difference in the percent of ITA7 colocalized with endothelial cells between KO and WT adipose tissue (p = 0.17). Values are mean ± standard error from unpaired t test with Welch's correction. Two tailed p value was calculated. n = 30 images from 4 WT animals and 25 images from 3 KO animals. (E,F) Representative masks of the images used to quantify the spatial proximity of ITA7⁺ cells with vessels of KO and WT adipose tissue. cytoNet analysis showed that (G) ITA7⁺ cells were closer to vessels in KO tissue compared to WT tissue (61.9 ± 14.6 pixels in KO vs. 96.5 ± 11.8 pixels in WT, p = 0.046), and (H) the number of ITA7⁺ cells was greater in WT tissue compared to KO, although this difference did not reach significance (6.8 ± 1.1 cells in KO vs 10.4 ± 1.1 cells in WT, p = 0.059).