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. Author manuscript; available in PMC: 2022 Sep 1.
Published in final edited form as: Hepatology. 2021 Jun 21;74(3):1560–1577. doi: 10.1002/hep.31831

Fig. 7. Notch1 signaling is required for the Hedgehog/SMO/Gli1 pathway-mediated Dvl2 activation and NEK7/NLRP3 inhibition in MSC-mediated immune regulation.

Fig. 7.

(A) BMMs were isolated from SMOFL/FL mice and co-cultured with MSCs or CD47-deficient MSCs followed by LPS stimulation. Immunofluorescence staining for Gli1 expression in macrophages (n=3-4 samples/group). DAPI was used to visualize nuclei. Scale bars, 40μm and 20μm. (B) Western blot analysis and relative density ratio of Gli1, Dvl2, NRX, β-catenin, NEK7, and NLRP3 in LPS-stimulated macrophages. (C) BMMs were isolated from SMOFL/FL mice and transfected with CRISPR/Cas9-Notch1 KO or control vector, and then co-cultured with MSCs followed by LPS stimulation. Immunofluorescence staining for Dvl2 expression in macrophages (n=3-4 samples/group). DAPI was used to visualize nuclei. Scale bars, 100μm and 40μm. (D) Western blot analysis and relative density ratio of NICD, Dvl2, NRX, β-catenin, NEK7, and NLRP3 in LPS-stimulated macrophages. (E) Caspase-1 activity (U) in macrophages after co-culture (n=3-4 samples/group). (F) qRT-PCR analysis of TNF-α, IL-1β, IL-6, CXCL-2, and CXCL-10 (n=3-4 samples/group). All Western blots represent three experiments, and the data represent the mean±SD. *p<0.05, **p<0.01, ***p<0.001.