B-cell differentiation associates with down-regulation of vimentin expression. B-cell differentiation associates with down-regulation of vimentin expression.

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B-cell differentiation associates with down-regulation of vimentin expression. B-cell differentiation associates with down-regulation of vimentin expression. (A, B) Representative flow cytometry plots with colour coding showing the decrease in vimentin level in CD138+ cells in vitro. Primary B cells were cultured in anti-IgM, CD40L, and IL-4 for 4 d to generate CD138+ plasma cells (A, left panels) and IgG1+ cells (A, right panels). On day 4 of the culture, the cells were labelled for surface expression of either CD138 or IgG1 before staining for intracellular vimentin. The samples were analysed using flow cytometry. The expression of vimentin in different populations was illustrated using (A) colour coding or (B) histogram. (C) Representative confocal images showing the distribution of intracellular vimentin in CD138− and CD138+ cells generated in vitro. Primary B cells were cultured in anti-IgM, CD40L, and IL-4 for 4 d to generate CD138+ plasma cells. On day 4 of the culture, the cells were labelled for surface expression of CD138 and fixed onto poly-lysine–coated coverslips. The cells were then permeabilised and labelled for vimentin before being imaged using confocal microscopy. Scale bar = 5 μm. (D, E) Representative flow cytometry plots showing the gating strategies for plasma cells generated in an in vivo setting and their corresponding vimentin expression compared with the naive counterparts. (D) WT mice were infected with 200 PFU influenza virus via intranasal inoculation. On day 9 after infection, the spleen (D) and the draining mediastinal lymph nodes (E) were harvested and single-cell suspensions were obtained. The cells were labelled for the surface expression of B220, CD19, IgD, and CD138, and then fixed and labelled for intracellular vimentin. The expression of vimentin in each population was characterised using flow cytometry. (F) Pooled data of the level of B and T cells’ reconstitution in μMT-KO recipients that received mixed BM containing WT or Vim−/− BM 6–8 weeks after adoptive transfer. t test was used to determine statistical significance. Each dot represents one mouse. Carlson Tsui et al. LSA 2018;1:e201800060 © 2018 Tsui et al.