Supplementary Figure 3: Cellular co-localization of reporter protein expression in the CNS and correlation with iNOS and arginase-1 expression in situ.
From: Mononuclear phagocytes locally specify and adapt their phenotype in a multiple sclerosis model
(a-d) Confocal images of the spinal cord of iNOS-tdTomato-cre (upper row; tdTomato, red) and (e-h) Arginase-YFP (lower row; YFP, green) mice perfused at peak of EAE and counterstained for marker proteins of phagocytes (a,e; Iba-1, gray), T cells (b,f; CD3, gray), oligodendrocytes (c,g; Olig2, gray) and astrocytes (d,h; GFAP, gray). Images are representative of 3 independent expeirments. Average percentage of co-localisation between tdTomato and Iba-1 ( ± s.e.m.) was 93.7 ± 2.5 (n = 3 mice and 447 cells analyzed); between YFP and Iba-1 ( ± s.e.m.) was 95.3 ± 1.5 (n = 3 mice and 601 cells analyzed). Scale bar 20 µm. (i) Confocal image of the spinal cord of an iNOS-tdTomato-cre mouse at weight loss (tdTomato, red; iNOS, gray). Scale bar, 20 µm. (j) tdTomato- and iNOS-fluorescence intensities in phagocytes analyzed in spinal EAE lesions of iNOS-tdTomato-cre mice at weight loss. iNOS was expressed in 98.6 ± 1.2% of all tdTomato+ cells (average ± s.e.m.; n = 5 mice and 732 cells analyzed). (k) Confocal image of the spinal cord of an Arginase-YFP mouse at remission (YFP, green; arginase-1, gray). Scale bar 20 µm. (l) YFP- and arginase-1-fluorescence intensities in phagocytes analyzed in spinal EAE lesions of Arginase-YFP mice at remission. Arginase-1 was expressed in 99.8 ± 0.2% of all YFP+ cells (average ± s.e.m.; n = 5 mice and 658 cells analyzed).
