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. 2021 Feb 3;11:2890. doi: 10.1038/s41598-021-82346-6

Figure 2.

Figure 2

NgR1 and NgR3 are localized to B-cells within lymphoid-like follicles of the spinal cord. (a) Western blot of spleen and spinal cord extracts from ngr1+/+ and ngr1−/− mice showcasing the expression of NgR1 and NgR3. β-actin was used as a loading control. (b) Semi-quantitative analysis of both NgR1- and NgR3-positive B-cells per unit area (mmy) of spleen and spinal cord infiltrates for the ngr1+/+ and ngr1−/− mice (n = 34 ngr1+/+ mice; 108 sections, n = 15 ngr1−/− mice; 40 sections) during the onset of EAE. These B-cells demonstrated a highly inducible receptor expression (student’s t test with Bonferroni correction ***p < 0.001 and **p < 0.01) compared to controls. (c) Illustration of inflammatory infiltrates analyzed via immunofluorescence immunolabeling of follicle-like structures (FLS) within the lumbosacral spinal cords of EAE-induced mice in (dk). (d) Representative NgR1 and B220 double-labeled images showing the expression on clusters of B-cells localized to infiltrates around the leptomeninges during the onset of EAE in ngr1+/+ mice. (e) Representative NgR3 and B220 double-labeled images showing the expression on clusters of B-cells localized to infiltrates around the leptomeninges during the onset of EAE in ngr1+/+ mice (hatched boxes). (f) Representative images for the ngr1−/− mice, highlighting the absence of the NgR1 protein expression in clusters of B-cells in similar infiltrates of the leptomeninges during the onset of EAE (hatched boxes). (g) Representative images for the ngr1−/− mice, highlighting the strong expression of the NgR3 protein expression in clusters of B-cells in similar infiltrates of the leptomeninges during the onset of EAE (hatched boxes). (h) Representative NgR1 and CD19 double-labeled images showing the expression on clusters of B-cells localized to infiltrates around the leptomeninges during the onset of EAE in ngr1+/+ mice. (i) Representative NgR3 and CD19 double-labeled images showing the expression on clusters of B-cells localized to infiltrates around the leptomeninges during the onset of EAE in ngr1+/+ mice. (j) Representative images for the ngr1−/− mice, highlighting the absence of the NgR1 protein expression in clusters of CD19 + B-cells in similar infiltrates of the leptomeninges during the onset of EAE (hatched boxes). (k) Representative images for the ngr1−/− mice, highlighting the strong expression of the NgR3 protein expression in clusters of CD19 + B-cells in similar infiltrates of the leptomeninges during the onset of EAE. Scale = 50 μm.