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. 2021 Oct 26;12:743354. doi: 10.3389/fimmu.2021.743354

Figure 5.

Figure 5

S100A9 expression increased in TMs and treatment with tasquinimod or LY294002 attenuated the ratio of M2-TMs following UPEC-induced orchitis. (A) Comparison of testis between the control and UPEC infection groups (8week, n=4/group). (B) The growth of the colonies after homogenization and plating of testis tissues in the control and UPEC groups (n=4/group). (C) Flow chart of preparation for TMs, testes from control and UPEC group were digested and interstitial cells were incubated with F4/80 magnetic beads (8week, n=4/group). TMs were obtained by magnetic cell sorting (MACS), and the purity was verified by flow cytometry for subsequent experiment. (D, E) Western blot analysis results and quantification data of S100A9 expression in TMs of control and UPEC group (8week, n=4/group). Data represent the mean ± standard error. **P < 0.01. (F) Immunohistochemistry staining result of S100A9 in the control and UPEC group testis (8week) at the 7th day after infection. Black arrows showed the expression of S100A9 (scale bar: 200, 50 μm). (G) Double immunofluorescence staining of S100A9 and mouse macrophage marker (F4/80) in the testis (8week) of control and UPEC group (scale bar: 200, 50 μm). (H) Flowcytometric analysis of total TMs (8week, n=2/group) was marked as CD45+F4/80+. Percentage of M2Φ in testis decreased with the treatment of S100A9 inhibitor (tasquinimod) or PI3K inhibitor (LY294002), compared with the UPEC group.