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. 2024 Aug 30;38(11):2355–2365. doi: 10.1038/s41375-024-02391-8

Fig. 3. IL-10 induces M2 polarization in TAMs through activation of STAT3 signaling.

Fig. 3

A Effect of IL-10 on macrophage M2 polarization in vitro. THP-1-derived and primary monocyte-derived macrophages (from 3 independent donors) were treated with 100 ng/ml rhIL-10 in the 3DTEBM for 3 days. Macrophage polarization was tested by flow cytometry and represented as M2/M1 ratio, and a fold-change of non-treated condition (NT). B Effect of IL-10 on STAT3 signaling in macrophages. THP-1-derived macrophages were treated with rhIL-10 (0, 10, 100 ng/ml) for 30 min, and STAT3 activation was measured by western blotting. C Effect of anti-IL-10R mAb on macrophage polarization in vitro. THP-1-derived and primary monocyte-derived macrophages (from 3 independent donors) treated with rhIL-10 (100 ng/ml) in combination with anti-IL-10R mAb (5 µg/ml) in the 3DTEBM for 3 days. Macrophage polarization was tested by flow cytometry and represented as M2/M1 ratio, and a fold-change of non-treated condition (NT). D Effect of anti-IL-10R inhibition on pSTAT3 activation in THP-1-derived macrophages. THP-1-derived macrophages were pretreated with or without anti-IL-10R mAb (5 µg/ml) for 30 min before addition of rhIL-10 (100 ng/ml), and STAT3 activation was measured by western blotting. E Effect of selective cellular STAT3 protein degradation on macrophage polarization. THP-1-derived macrophages were treated with the STAT3 proteolysis targeting chimera (PROTAC) molecule SD-36 (2.5uM) for 24 h. Then, rh-IL-10 (100 ng/ml) was added for 3 additional days. Macrophage polarization was tested by flow cytometry and represented as M2/M1 ratio, and a fold-change of non-treated condition (NT). (Bars = Average ± SD, **p < 0.01).