Fig. 3: SGK1 co-localizes and interacts with NLRP3. | Nature Communications

Fig. 3: SGK1 co-localizes and interacts with NLRP3.

From: Serum/glucocorticoid-inducible kinase 1 deficiency induces NLRP3 inflammasome activation and autoinflammation of macrophages in a murine endolymphatic hydrops model

Fig. 3

a Immunofluorescence staining for DAPI (blue), NLRP3 (green), and SGK1 (red) in THP-1 cells that were stimulated with LPS (200 ng/mL, 3 h) and ATP (5 mM, 30 min), scale bar = 50 μm. b Representative confocal images of HEK-293T cells transiently expressing GFP-NLRP3 (green), FLAG-SGK1 (red), MYC-ASC (magenta), and staining DAPI (blue), which were stimulated with LPS (200 ng/mL, 3 h) and ATP (5 mM, 30 min), scale bar = 50 μm. c Immunofluorescence staining for DAPI (blue), NLRP3 (green), CD68 (red), and SGK1 (magenta) in primary mVRML cells of WT mice which were stimulated with LPS (200 ng/mL, 3 h) and ATP (5 mM, 30 min), scale bar = 10 μm. d Immunofluorescence staining for DAPI (blue), NLRP3 (green), IBA1 (red), and SGK1(magenta) in macula of MD patients, scale bar = 10 μm. e Co-IP analysis of the interaction of NLRP3 and SGK1 from the total lysates of THP-1 cells which were stimulated with LPS (200 ng/mL, 3 h) and ATP (5 mM, 30 min). f Co-IP analysis of the interaction between GFP-NLRP3 and FLAG-SGK1 from the total lysates of HEK-293T cells which were stimulated with LPS (200 ng/mL, 3 h) and ATP (5 mM, 30 min). g GST pull-down assay with GST-fused NLRP3 and in vitro translated SGK1. h Co-IP analysis of the interaction of NLRP3 and SGK1 from the total lysates of THP-1 cells which were stimulated with saline, LPS (200 ng/mL, 3 h) or LPS + ATP (5 mM, 30 min). i Domain-mapping experiment showing details of the NLRP3-SGK1 interaction in plasmid-transfected HEK-293T cells. mVRMLs, mouse vestibular-resident macrophage-like cells; Casp-1 caspase-1. Source data are provided as a Source Data file.

Back to article page