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

Figure 4.

Figure 4

The triggering receptors on microglial cells 2 (TREM2) interact with Jak3 in the brain, and the intestinal deficiency of Jak3 leads to reduced interactions and expression of TREM-2 in the brain with Jak3 during HFD-induced obesity: (A) Jak3 interactions with TREM-2 were determined using brain tissue lysates from ND- and HFD-fed mice. Lysates from WT (littermate control), Jak3-KO, IEC-Jak3-KO and flox-Jak3 (littermate control) were subjected to IP followed by IB using the indicated antibodies. Blots shown represent n = 3 experiments. (B) The impact of global and intestinal epithelial tissue-specific deficiencies of Jak3 on the brain expression of β-Amyloid receptor TREM2 on microglial cells was determined in ND- and HFD-fed mouse brain. Representative flow cytometric histogram graphs of individual mouse brain cells showing the microglial levels of expression of TREM-2 receptor in the four groups (n = 5/group; WT-ND (littermate control), Jak3-KO-ND, WT-HFD (littermate control), Jak3-KO-HFD) are shown in the left panel, and the corresponding histogram bar graphs indicating mean ± SD values are shown for the comparative average cell counts for the indicated groups of mice. * Indicate statistically significant difference from the corresponding controls (KO-ND p = 0.05, KO-HFD p = 0.03). ** Comparison between Int-KO ND and Int-KO HFD group. (C) Brain tissue sections from WT-control littermate and Jak3-KO mice fed with either ND or HFD were immunostained using TREM-2 primary antibodies followed by FITC secondary antibodies, and mounting media containing PI were used to visualize the nucleus. Representative images (n = 10) are shown. (D) Data from similar experiments as in “B” but for the four different groups of mice (n = 5/group; flox-Jak3-ND (littermate control), IEC-Jak3-KO-ND, flox-Jak3-HFD (littermate control), IEC-Jak3-KO-HFD) are shown. * Indicate statistically significant difference from the corresponding controls (IEC-Jak3-KO-ND p = 0.05, KO-HFD p = 0.05). (E) Brain tissue sections from flox-Jak3-control littermate and IEC-Jak3-KO mice fed with either ND or HFD were immunostained using TREM-2 primary antibodies followed by FITC secondary antibodies, and mounting media containing PI were used to visualize the nucleus. Representative images (n = 10) are shown.