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Fig. 4 | Arthritis Research & Therapy

Fig. 4

From: Semaphorin 3G exacerbates joint inflammation through the accumulation and proliferation of macrophages in the synovium

Fig. 4

Enhanced macrophage proliferation by Sema3G. A Chemotaxis assay. LPS-stimulated BMMs were subjected to a transwell migration assay. PBS (negative control), MCP1 (positive control), or various concentrations of Sema3G were added to the lower chambers. Cell migration towered to the lower chamber was determined by Hoechst staining. The pictures were taken for five different areas, and Hoechst-positive round cell numbers were counted. N = 7 from three independent experiments. One-way ANOVA test, followed by Dunnet test, was used for the statical analysis. B The volcano plot of Sema3G-stimulated BMMs. Genes differentially expressed are plotted as red dots (upregulated by Sema3G) or blue dots (downregulated by Sema3G). Some immune-related gene names are labeled. C EdU uptake in Sema3G-stimulated BMMs. Sema3G-stimulated or control (PBS) BMMs were pulsed with 10 μM EdU for 2 h, and EdU and Hoechst were detected by immunofluorescent analysis. The pictures were taken for ten different areas, and the percentage of EdU-positive cells over Hoechst-positive cells was calculated. Data were obtained from three independent experiments. N = 6. D The clinical score of Sema3G- or PBS-injected paws during CAIA. WT mice were subjected to CAIA. Sema3G (100 ng) was injected into the right footpad, and PBS was injected into the left footpad daily from day 3 to day 9. N = 11 from two independent experiments. E Infiltrating cell subsets to each paw on day 9 of CAIA. The hind paws were digested, and cells recovered were analyzed by flow cytometry. The percentages of T cells, B cells, and macrophages are shown. A paired t-test was used for the statistical analyses

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