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. 2022 Sep 9;14(18):3715. doi: 10.3390/nu14183715

Figure 2.

Figure 2

Intestinal epithelial cell (IEC) deficiency of Jak3 predisposes to exaggerated symptoms of HFD-induced obesity and glycemic dysregulation. (A), Genetic manipulation shows IEC deficiency of Jak3. Colonic tissue sections from floxed Jak3 (jak3f/f) or IEC-Jak3-KO (vil1-Cre- jak3f/f) were co-immunostained with IEC marker villin (green) and Jak3 (red) antibodies using DAPI as control for the nucleus. Representative images (n = 10) are shown from each group (n = 6). (B), Western blot analyses were performed using tissue lysates from floxed Jak3 and IEC-Jak3-KO mice for Jak3 expression (upper panel) using β-actin as control (lower panel). Representative blots (n = 3) are shown. (C), Fluorescence-activated cell sorting (FACS) analysis is presented as four quadrant dot plots to determine the impact of HFD on IEC-Jak3 expression by determining the double positive cells (Quadrant:UR) for IEC marker villin and Jak3 under ND or HFD. Jak3 positive cells are shown on the x axis and those of villin positive cells on the y axis. (D), Bar chart results following FACS experiments in “C” were repeated (n = 5), and mean ± SD values are shown as bar graph for the comparative average cell counts for the indicated groups of mice. *** denotes p = 0.004 (Int-KO ND), p = 0.008 (FL HFD), p = 0.001(Int-KO HFD). (E), IEC deficiency of Jak3 predisposes to increased body weight. Vil1-Cre/ jak3f/f (Int-KO) or their littermate controls jak3f/f (FL) mice were cohoused and subjected to either ND or HFD. Body weights of co-housed Int-KO and their FL littermate males were plotted at the indicated intervals as a percentage of their body weight at the time of weaning (n = 5/group). *** denotes p = 0.005 (Int-KO HFD), p = 0.05 (Int-KO ND). (F), IEC deficiency of Jak3 impairs the ability to restore blood glucose. Blood glucose concentrations in fasted mice were measured after intraperitoneal injection with 2 g/kg body weight of d-glucose followed by measuring the level of blood glucose at thew, indicating intervals post-injection (n = 5/group). *** denotes p = 0.003 (Int-KO HFD), p = 0.05 (Int-KO ND).