*2.5. GH Mitigates White Adipose Tissue Browning in CKD Mice*

Beige adipocyte cell surface markers' (CD137, Tbx1, Tmem26, Prdm16, Pgc1α, and Cidea) mRNA expression in inguinal WAT was normalized or decreased in GH-treated CKD mice relative to vehicle-treated CKD mice (Figure 4A–F). In WAT, de novo browning recruitment is promoted by the activation of Cox2/Pgf2α pathway and toll-like receptor Tlr2 and adaptor molecules, such as Myd88 and Traf6 [22]. GH treatment normalized expression of inguinal WAT Cox2, Pgf2α, Tlr2, Myd88, and Traf6 in CKD mice (Figure 4G–K).

**Figure 3.** Lipolytic gene expression and protein content in CKD mice. CKD mice were fed ad libitum, whereas other mouse groups received an energy intake amount equal to that of CKD + Vehicle mice. By using qPCR, the expression of lipolytic genes (Atgl and Hsl) in the inguinal WAT was determined (**A**,**C**). In addition, the total protein content of ATGL and HSL as well as the relative phosphorylated HSL/total HSL ratio in the inguinal WAT were evaluated (**B**,**D**,**E**). Results are analyzed and expressed as in Figure 2. Data are expressed as mean ± SEM. For comparison of the means between two groups, data were analyzed by Student's 2-tailed *t*-test. Posthoc analysis was performed with Tukey's test. Specific *p*-values are shown above the bar. ns signifies not significant, \* *p* < 0.05, \*\* *p* < 0.01.

**Figure 4.** GH reduces browning of adipose tissue in CKD mice. CKD mice were fed ad libitum, whereas other mouse groups received an energy intake amount equal to that of CKD + Vehicle mice. qPCR was used to assess the gene expression of the beige adipocyte markers CD137, Tbx-1, Tmem26, Prdm16, Pgc1α, and Cidea in the inguinal WAT (**A**–**F**). In addition, inguinal WAT was also used to evaluate the gene expression of the Cox2 signaling pathway and the toll-like receptor pathway (Cox2, Pgf2, Tlr2, Myd88, and Traf6) (**G**–**K**). Final results were expressed in arbitrary units, with one unit being the mean level in Sham + Vehicle mice. Results are analyzed and expressed as in Figure 2. Data are expressed as mean ± SEM. For comparison of the means between two groups, data were analyzed by Student's 2-tailed *t*-test. Posthoc analysis was performed with Tukey's test. Specific *p*-values are shown above the bar. ns signifies not significant, \* *p* < 0.05, \*\* *p* < 0.01.
