Fig 1: IL-33 induces the morphologic change of oligodendrocytes. a Mature OLGs were exposed to 10 ng/ml of IL-33 for 24 h, and then subjected to immunofluorescence (red) using anti-MBP in accompany with DAPI nuclear counterstaining (blue). The interconnected network of OLG processes along with a small cell shape was injured after exposure to IL-33 (arrows). The complex network shape of OLGs in the control culture is indicated by arrowheads. In addition, MBP+-OLGs in the cultures treated with vehicle and IL-33 were quantified. The cell size of MBP+-OLGs was measured using NIH ImageJ analysis software. b Total proteins were prepared from the cultures treated with vehicle and IL-33. The samples were subjected to western blot analysis for the measurement of MBP. Data are presented as mean ± SEM of the three independent experiments. **p < 0.01, ***p < 0.001 versus the vehicle group. Scale bar in a 50 μm
Fig 2: IL-33 in hypothalamus was increased by chronic HFD feeding. a The brain sections were prepared from adult mice. The brain sections containing hypothalamic regions were subjected to double immunofluorescence for IL-33 (green) with various glial markers (red) including Iba1 (microglia), GFAP (astrocytes), and Olig2 (OLGs). The nuclear counterstaining by DAPI (blue) was also conducted. The representative images were taken from hypothalamic ARC. The enlarged images from the insets indicated in the upper panel are included in the lower panel. Arrows shown in the lower panel indicate IL-33+/GFAP+-astrocytes and IL-33+/Olig2+ OLGs. b Total proteins were prepared from hypothalamus of the obese mice at 3 and 4 month after Chow or HFD feeding, and then subjected to western blot analysis (left-hand panel) to examine IL-33 protein expression. IL-33 protein levels were quantified (right-hand panel). c, d The brain tissue sections were prepared from the Chow- and HFD-fed groups at 4 month after feeding, and then subjected to immunofluorescence. The representative images were captured from ARC regions. Arrows indicate IL-33+/GFAP+-astrocytes (c) or IL-33+/Olig2+-OLGs (d). The results shown in b are presented as mean ± SEM (n = 3 animals for each group at each time point). *p < 0.05, ***p < 0.001 versus the Chow group. Scale bar in a, d 50 μm; in c 100 μm
Supplier Page from Enzo Life Sciences, Inc. for IL-33 (human), (recombinant)