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Fig. 6 | Acta Neuropathologica Communications

Fig. 6

From: Dysregulation of lysophosphatidic acids in multiple sclerosis and autoimmune encephalomyelitis

Fig. 6

EAE in LPAR2 deficient mice. a Time course of the clinical EAE scores and body weight of LPAR2−/− versus LPAR2+/+ mice in the MOG-induced primary progressive EAE model (n = 19 for LPAR2−/− and n = 15 for LPAR2+/+). LPAR2 mice are on a mixed C57BL6xSv129 genetic background (Sv129 EAE unresponsive), so that the wild type LPAR2+/+ mice have very mild signs of EAE. AUCs of the scores and body weights were compared with Mann Whitney U test and 2-tailed unpaired t-test, respectively. P < 0.05. b Box plot of the sum of all LPAs in the spleen and autotaxin in serum 36 days after immunization. The boxes show the interquartile range, the line is the median, the whiskers show minimum to maximum and the dots are results of individual mice. The asterisk shows a statistically significant difference between groups, 2-tailed unpaired t-test, P < 0.05. c Exemplary immunofluorescence images taken from the lumbar spinal cord of LPAR2−/− and LPAR2+/+ mice. The images show the myeloid marker, Iba-1 with a neuronal counterstain of NeuN and DAPI for labeling of nuclei. The dashed rectangles show the areas taken for zoom-in images. Scale bar 50 μm and 25 μm in zoom-in images. Cell counts for Iba-1 or CD11b positive cells were obtained with FIJI ImageJ for n = 6–9 images of 3–4 mice per group and compared with a 2-way ANOVA for “marker” versus “genotype”. The counts differed significantly for Iba-1 between genotypes (*P < 0.05), and the morphology of large rhomboid microglia in LPAR2−/− mice suggested a stronger activation. d FACS analysis of the number of CD4+ and CD8+ T-cells in the lumbar spinal cord in LPAR2−/− and LPAR2+/+ mice 36 days after immunization. The asterisks show significant differences between LPAR2−/− and LPAR2+/+ mice (2-way ANOVA and subsequent t-tests according to Dunnett for each cell population, P < 0.05). e Exemplary forward (FSC) versus sideward (SSC) scatter plots of the immune cells in the spleen in LPAR2−/− and LPAR2+/+ mice 36 days after immunization. Lymphocytes, monocytes and DCs (myeloid cells) were identified by size and granularity. f Box plots showing the quantification of lymphocytes, T-cell subtypes and B-cells in the spleen in LPAR2−/− and LPAR2+/+ mice 36 days after immunization (meaning of boxes as in b). Asterisks denote significant difference between groups (* P < 0.05, *** P < 0.001, one-way ANOVAs with subsequent t-tests versus EAE-LPAR2+/+ mice according to Dunnett). g, h Box plots showing the quantification of myeloid cells in the spleen in LPAR2−/− and LPAR2+/+ mice 36 days after immunization. Expression of the homing receptor CCR7 by dendritic cells is a marker of their activation (meaning of boxes as in b, statistics as in f)

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