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. 2022 Apr 7;14(2):245–270. doi: 10.1016/j.jcmgh.2022.04.002

Figure 8.

Figure 8

KO of TM9SF4 promoted M1 macrophage polarization but inhibited M2 macrophage polarization. (A and B) Colonic CD11b+ cells from 2% DSS-challenged WT or KO mice were purified by magnetic-activated cell sorting. The mRNA levels of (A) M1 macrophages markers IL23p19, IL6, iNOS, and (B) M2 macrophages markers MRC1, Ym1, Arg1 were assessed by qRT-PCR (n = 4). Representative images and data summary for immunohistochemical staining of (C) CD80 and (D) CD206 in the colon tissues of DSS-treated WT/KO mice (n = 7). Brown, CD80/CD206-positive signals; blue, nuclear counterstain. Scale bars: 100 μm. (E) Expression levels of TM9SF4 in BMDMs and Raw264.7 macrophages were compared by Western blot (n = 3). BMDMs were treated with LPS/IFNγ or vehicle, followed by (F) qRT-PCR–based mRNA level analysis of IL23p19, IL6, IL12a, and iNOS (n = 4–6) or by (H) FACS analysis of CD11c+ M1 BMDMs (n = 4). (G and I) BMDMs were treated with IL4/IL13 or vehicle, followed by (G) qRT-PCR–based mRNA level analysis of Mrc1, Ym1, Il10, and Arg1 (n = 4–6) or by (I) FACS analysis of CD206+ M2 BMDMs (n = 4). (J) BMDMs were challenged with LPS/IFNγ. The levels of GRP78, ATF4, cleaved ATF6, spliced XBP1, CHOP, IL1β, and iNOS were detected by Western blot (n = 3). (K) Enhanced aggresome formation in BMDMs from KO mice by flow cytometry analysis (n = 5). Means ± SEM. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; MFI, mean fluorescent intensity.