Extended Data Fig. 2: Rspondin3 regulates macrophage phenotype transition via an angiocrine manner. | Nature Immunology

Extended Data Fig. 2: Rspondin3 regulates macrophage phenotype transition via an angiocrine manner.

From: The angiocrine Rspondin3 instructs interstitial macrophage transition via metabolic–epigenetic reprogramming and resolves inflammatory injury

Extended Data Fig. 2

a, Levels of anti-inflammatory markers (CD206, CD301, arginase 1, IL-10) and pro-inflammatory markers (CD86, CD80, TNF, iNOS) measured by flow cytometry in BMDMs incubated with the EC conditioned medium collected from lung ECs isolated from wildtype mice (WT EC), Rspo3EC-/- mice (ECRspo3-/-) at baseline or endotoxemia conditions (LPS i.p., 12 mg/kg for 24h) (LPS/WT EC, LPS/ ECRspo3-/-), respectively; Data are representative of three independent experiments with n=3 samples per group (mean ± sd), statistical significance was determined by two-way ANOVA with Tukey’s multiple comparisons test using GraphPad Prism with individual P values (left to right) are: CD206 (****P<0.0001, ns P=0.9405, ***P=0.0002, ****P<0.0001), CD301 (****P<0.0001, ns P=0.9991, ****P<0.0001, ****P<0.0001), arginase1 (****P<0.0001, ns P=0.4164, ****P<0.0001, ****P<0.0001), IL-10 (****P<0.0001, ns P=0.9933, ****P<0.0001, ****P<0.0001), CD86 (****P<0.0001, ns P=0.8224, ***P=0.0001, ns P= 0.9075), CD80 (****P<0.0001, ****P<0.0001, ns P=0.7688), TNF (****P<0.0001, ns P=0.4519, **P=0.0061, ns P=0.9997), iNOS (****P<0.0001, ns P=0.9996, ns P=0.6726, ns P>0.9999). b, Levels of anti-inflammatory markers (CD206, CD301, arginase 1, IL-10) and pro-inflammatory markers (CD86, CD80, TNF, iNOS) measured by flow cytometry in BMDMs co-cultured in-contact with the ECs from wildtype mice (WT EC), or Rspo3EC-/- mice (ECRspo3-/-) at basal conditions or mice challenged with sublethal LPS (WT EC/LPS, ECRspo3-/-/LPS), respectively; Data are representative of three independent experiments with n=3 samples per group (mean ± s.d), with each dot representing an individual sample. Statistical significance was determined by two-way ANOVA with Tukey’s multiple comparisons test using GraphPad Prism with individual P values (left to right) are CD206 (****P<0.0001, ns P>0.9999, ns P>0.9999), CD301 (****P<0.0001, ns P=0.9977, ns P=0.9998), arginase1 (****P<0.0001, ns P=0.9328, ns P=0.9998), IL-10 (****P<0.0001, ns P=0.8989, ns P=0.9393), CD86 (ns P=0.8308, ****P<0.0001, ns P=0.9600), CD80 (ns P>0.9999, ****P<0.0001, ns P=0.9679), TNF (ns P=0.9997, ****P<0.0001, ns P=0.9903), iNOS (ns P= 0.9996, ****P<0.0001, ns P= 0.2748).

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