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. 2019 Oct 2;9:14189. doi: 10.1038/s41598-019-50591-5

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Development of myeloma with associated spinal glial cell activation. (a) Serum IgG2b levels were quantified by ELISA throughout the experiment. Statistical comparison by two-way ANOVA with Sidak’s multiple comparisons test. (b) Flow cytometry analysis of GFP+ (5TGM1) population in bone marrow and spleens, displayed as percentage of total cell count. Statistical comparisons by t-test with Holm-Sidak multiple comparisons correction. (c) Representative images showing GFAP (red channel) staining in dorsal horn region (indicated by dashed line) of T8 spinal cords from non-tumour and 5TGM1 MM-bearing KaLwRij mice, with d) normalised integrated density of GFAP staining. (e) Representative images of Iba1 staining (red channel) in the same region with Iba1+ cell bodies indicated with arrowheads, and (f) cell counts per dorsal horn for non-tumour and MM-bearing mice. All histology images counterstained with DAPI (blue channel), and white scale bars represent 100 µm. Statistical comparisons in (d) and (f) by unpaired Student’s t-test. All statistical comparisons: ns: not significant, *p < 0.05; **p < 0.01; ***p < 0.001.