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. 2022 Nov 3;13:6320. doi: 10.1038/s41467-022-33796-7

Fig. 4. VWF triggers M1 macrophage phenotype.

Fig. 4

a–d Murine BMDMs were incubated in the presence or absence of a variety of different agonist combinations including (LPS 100 ng/ml and IFN-γ 20 ng/ml), (IL-4 40 ng/ml, IL-10 10 ng/ml, and IL-13 20 ng/ml) or pd-VWF (10 µg/ml) for 24 h and then cell surface marker expression was examined by flow cytometry. Flow gating strategy is presented in Supplementary Fig. 15. a Untreated control BMDMs expressed no CD38 or CD206. b The majority of BMDMS treated with LPS and INF-γ were CD38 positive, consistent with an M1 phenotype (*P = 0.0004). c In contrast, the majority of BMDMs incubated with IL-4, IL-10, and IL-13 were CD206 positive, consistent with an M2 phenotype (*P < 0.0001). d BMDM stimulation with pd-VWF (10 μg/ml) resulted in a significant increase in expression of CD38, consistent with a pro-inflammatory M1 macrophage phenotype (*P < 0.0001). Consistent with this M1 phenotype, e VWF treatment also resulted in a significant increase in iNOS expression in BMDMs (*P = 0.0287), f together with a marked increase in generation of reactive oxygen species (ROS). The data are presented as mean values ± SD for three independent experiments. The significance was calculated by ANOVA where *P < 0.05, **P < 0.01, ***P < 0.001 ****P < 0.0001; ns = not significant. Source data for this figure are provided as a Source Data file.