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. 2022 May 3;13:2406. doi: 10.1038/s41467-022-30174-1

Fig. 2. T-cell-specific Mincle deletion protects mice from EAE.

Fig. 2

a Targeting vector design for the generation of a mouse strain with flanking Clec4e exon 3-5 by loxP sites and western analysis of Mincle protein expression in TH17 cells and bone marrow macrophages (1 μg/ml LPS, 6 h) from Mincle f/+Lck-Cre and Mincle f/fLck-Cre mice, n = 2 for each genotype, density values measured using Image J for the representative blot shown, ND not detected. b Mean clinical score of EAE in Mincle f/+Lck-Cre and Mincle f/fLck-Cre mice (n = 6 mice in each group) induced by active immunization with MOG35-55. c, d Absolute cell numbers (c) and gating strategy (d) of CNS-infiltrating cells were measured at the peak of disease by analyzing brain mononuclear infiltrating cells through flow cytometry with indicated antibodies, n = 4 biological replicates. e Real-time PCR analysis of relative mRNA expression of inflammatory genes in the spinal cord from Mincle f/+Lck-Cre and Mincle f/fLck-Cre mice at the peak of disease. Expression was normalized to β-actin mRNA, n = 4 biological replicates. f Hematoxylin and eosin (H&E) staining (upper panels) and Luxol fast blue staining (lower panels) of lumbar spinal cords from Mincle f/+Lck-Cre and Mincle f/fLck-Cre mice harvested at the peak of disease, Scale bars represent 100 μm. Arrows in the upper panel indicate inflammatory cells infiltration, and arrows in the lower panel indicate demyelination area. Representative data are shown for n = 4. g Flow cytometry analysis of infiltrated cytokine-producing CD4 T cells in CNS at the peak of disease, n = 4 biological replicates. *P < 0.05, **P < 0.01 (Two-sided student’s t test, c, e). *P < 0.05 (Two-way ANOVA for b). Data are represented as mean ± SD. Exact P values for asterisks (from left to right): b 0.0002 c 0.0032 0.0012 0.0017 0.0068 d 0.0446 0.0053 0.0062 0.0341 0.0410 0.0021 0.0023 0.0003 0.0005 0.0001 g 0.0014 0.0030.