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Fig. 2 | Molecular Medicine

Fig. 2

From: The involvement of neuroimmune cells in adipose innervation

Fig. 2

LysMCre±::BDNF−/− (KO) have increased adiposity and impaired response to cold due to genetic denervation of scWAT. Adult (22–23 week old) male LysMCre−/−::BDNFfl/fl (CON) and LysMCre±::BDNF−/− (KO) mice were cold exposed at 5 °C for 4 days; body weight and adiposity were compared between CON and KO groups, p = 0.0750 and p = 0.1107, respectively (a). Body and tissue weight data were analyzed by two-tailed Student’s T-Test, N = 5 CON, N = 6 KO. Protein expression of PGP9.5 and tyrosine hydroxylase (TH) in inguinal scWAT was measured by Western blotting from adult (12–25 week old) 7-day cold (5 °C) exposed WT/CON and KO male animals, p = 0.0025 and p = 0.0200, respectively (b). β-Tubulin was used as a loading control for normalization. Data were analyzed by two-tailed Student’s T-Test, N = 3 WT/CON, N = 4 KO, *denotes data that was removed from analysis due to lack of expression of loading control. Gene expression of Ucp1 was measured in adult (12–25 week old) 7-day cold (5 °C) exposed WT/CON and KO males, p = 0.0573 (c). Data were analyzed by two-tailed Student’s T-Test, N = 5 WT/CON, N = 5 KO. Circulating thyroid hormones, triiodothyronine (T3), and thyroxine (T4) were measured by ELISA from serum of adult (22–23 week old) 4-day cold (5 °C) exposed CON and KO male mice, p = 0.9526 and p = 0.3471, respectively (d). Data were analyzed by One-way ANOVA, with Tukey’s multiple comparisons test, N = 5 CON, N = 6 KO. Immunofluorescent staining for UCP1 was performed on inguinal scWAT sections of adult (22–23 week old) male CON and KO mice following 4-day cold (5 °C) exposure (e). Immunofluorescent staining for UCP1 and Perilipin was performed on inguinal scWAT sections of adult (22–23 week old) male KO mice following 4-day cold (5 °C) exposure (f). Typogen Black, used to quench lipid autofluorescence, provided staining cell morphology which was visualized under brightfield microscopy. Overlay is immunofluorescence over brightfield of the same area (f). Images were acquired with a 10X or 40X objective and are representative of N = 5 CON, N = 8 KO (E) and N = 4 KO (f). For all error bars are SEMs

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