Fig. 5.

Expression and regulation of LPARs. a, b FACS analyses of splenic immune cells in control and EAE mice showing exemplary scatter plots of T-helper and T-suppressor cells gated according to CD4+ versus LPAR2 and CD8+ versus LPAR2. The spleens were taken during the second interval in SJL/J mice, 35 days after immunization. Control mice had received an injection of CFA without PLP. The green framed area shows the double positive cells, which were used for quantification. c Exemplary histograms of myeloid cells gated according to the expression of LPAR2. Each 3 examples of control and EAE mice are shown. d, e, f Box and scatter plots of the total numbers of LPAR positive CD4+ and CD8+ T-cells and of CD11c ∩ F4/80 positive myeloid cells. 100,000 cells were counted of each 6 mice. Boxes show the interquartile range, whiskers minimum to maximum and the circles show results of individual mice. The line is the median. Data were compared with unpaired 2-tailed t-tests for each LPAR separately. *P < 0.05, **P < 0.01. g Box and scatter plots of the mRNA levels of LPAR1, 2, 3, 4 and 5 in the spleen, white blood cells (WBCs) and spinal cord in SJL-EAE mice, 35 days after immunization i.e. after the second peak. The mRNA levels were normalized to the delta Ct level of LPAR1 in control mice, which was set to 100%. Hence, the data show EAE effects and overall differences in the tissue specific LPAR expression patterns. Data were compared with unpaired 2-tailed t-tests for each LPAR separately. *P < 0.05, **P < 0.01, *** P < 0.001, meaning of the boxes as in (d–f)