Fig. 7.

Decreased claudin-1 immunoreactivity in the perineurium and vessel-associated macrophages in prolonged STZ DPN. Sciatic nerve from male Wistar rats was stained with anti-claudin-1 (a, b), anti-claudin-5 + anti-von Willebrand factor (vWF, c, d). Claudin-5 was quantified relative to β-actin by Western blot (e). Vessel-associated macrophages were identified by immunolabeling with anti-CD68 and anti-vWF (f, g) or anti-CD206 and RECA-1 (h, i). Fluorescence intensity was quantified in the regions of interest (perineurium; a, b) and (vWF+ vessels; c, d) normalized to the area. f, g, h, i The total number of macrophages (CD68+ or CD206+) in proximity to blood vessels (vWF+ or RECA-1+) was counted and expressed relative to the total number of capillaries in the endoneurium. n = 5–6. Arrows depict examples. Scale bars = 100 μm and in the insert 20 μm (f, g) and 10 μm (h, i), dot plots represent medians and interquartile range, Mann-Whitney test for comparison. *P < 0.05 and **P < 0.01