LMW Heparin Prevents Increased Kidney Expression of Proinflammatory Mediators in (NZBxNZW)F1 Mice
Figure 3
Characterization of nucleosomes used in this study and spontaneous splenocyte proliferation. Immunohistochemistry analysis detecting CCR2, IL1β, and TLR7 protein expression in kidneys of 31 w.o heparin or untreated B/W mice (a). SDS PAGE of nucleosomes and HMGB1 used in cell culture stimulations ((b), lane 1: MW standard, lane 2: nucleosomes (3 μg measured as DNA), and lane 3: HMGB1 (1 μg)). Western blot showing HMGB1 (arrows) in nucleosomes purified from A31 cell line ((c), lane 1: MW standard, lane 2: HMGB1 (0.1 μg), and lane 3: nucleosomes (2 μg measured as DNA)). Agarose gel electrophoresis of nucleosomes showing a distribution of nucleosome sizes ((d), lane 1: MW standard, lane 2: nucleosomes (3 μg measured as DNA)). Stimulation index (SI) of cells from prenephritic mice (e) and nephritic mice (f) stimulated with conA with or without heparin. Splenocytes from nephritic mice have a significantly higher proliferation (counts per minute, CPM) at 20 hours which persisted over time compared to splenocytes from prenephritic mice (g). Mean values and SD were calculated from triplicates from each cell culture. *Statistically significant () compared to medium stimulated cells at the same time point. Scale = 100 μm.
