Fig. 1: Decrease in SGK1 expression is associated with inflammasome activation in VRML cells from MD patients. | Nature Communications

Fig. 1: Decrease in SGK1 expression is associated with inflammasome activation in VRML cells from MD patients.

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

Fig. 1

a Representative confocal microscopy images showing IBA1 (red) and DAPI (blue) staining in the ampulla and macula of AN (n = 5) and MD patients (n = 5). Analysis of images is shown on the right, quantifying the number of IBA1-positive cells per area. The right panels represent 2-fold enlargements of the white-boxed areas in the left panels. Scale bar = 25 μm. b Representative images of immunohistochemical staining and densitometric quantification of IL-1β, IL-18, and Casp-1 in the ampulla and macula of AN (n = 4) and MD patients (n = 4), scale bar = 100 μm. c Quantitative real-time PCR analysis of IL1B, IL18, and CASP1 mRNA in VEO of AN (n = 9) and MD patients (n = 9). d ELISA of plasma IL-1β, IL-18, and Casp-1 levels in controls (n = 20) and MD patients (n = 20). e Quantitative real-time PCR analysis of SGK1 mRNA in VEO of AN (n = 9) and MD patients (n = 9). f Quantitative real-time PCR analysis of SGK1 mRNA in VRML cells of AN and MD patients (n = 3). g Representative images of western blotting detecting NLRP3 and SGK1 in VRML cells of AN and MD patients (n = 3). h Representative confocal microscopy images showing IL-1β (green), SGK1 (red), IBA1 (gray), and DAPI (blue) staining in the ampulla and macula of AN and MD patients, scale bar = 10 μm. i Immunofluorescence staining for Sox2 (green) and Myosin VIIa (red) in basilar membrane of neonatal mice in vitro, after treatment with IL-1β (10 ng/mL, 24 h). White asterisks indicate inner hair cell loss. Scale bar = 50 μm. j Immunofluorescence staining for TUNEL (green) and Myosin VIIa (red) in the basilar membrane at P3, after treatment with IL-1β (10 ng/mL, 24 h), scale bar = 50 μm. In each box plot, the central mark indicates the median, the bottom and top edge of the box indicate the interquartile range, and the whiskers represent the max and minimum data point. Results are presented as mean ± SD. Statistical analyses were carried out via two-sided t-test for a–f. VEO vestibular end organs, hVRMLs human vestibular-resident macrophage-like cells, Casp-1 caspase-1. Source data are provided as a Source Data file.

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