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. 2022 Aug 22;7(16):e158238. doi: 10.1172/jci.insight.158238

Figure 6. Nlrp3 deficiency protects against preterm birth by limiting decidual inflammation.

Figure 6

(A) Pregnant Nlrp3+/+ and Nlrp3–/– mice were intra-amniotically injected with LPS (100 ng/25 μL) or PBS on 16.5 days post coitum (dpc). We collected the decidua basalis on 17.5 dpc. The spatial localization of the murine decidua is shown. (B) Heatmap visualization of inflammatory gene expression in the decidua (n = 13–15 per group). (C) Expression of the inflammation-associated genes (n = 13–15 per group). (D) Immunoblotting of pro-CASP and CASP-1 p20 in the decidua (n = 6 per group). The expression was normalized by ACTB and shown as relative quantification. (E) Concentrations of total IL-1β in the decidua. Values were adjusted by the total protein concentration in each sample (n = 8–9 per group). (F) Immunoblotting of mature IL-1β in decidual tissue lysates immunoprecipitated with anti–IL-1β antibody (n = 3 per group). (G) Representative immunofluorescence images showing the expression of neutrophil elastase (NE) in the decidua (n = 5–7 per group). All images were taken at 200× original magnification. Scale bars represent 20 μm. Inset images show isotype control staining from the same case. (H) Immunoblotting of 2-chain NE in decidual tissue lysates (n = 6 per group). Data are shown as box-and-whisker plots where midlines indicate medians, boxes indicate interquartile ranges, and whiskers indicate minimum and maximum values. The P values of the comparisons between PBS- and LPS-injected dams of each genotype were determined by Mann-Whitney U test. *P < 0.05; **P < 0.01; ***P < 0.001; (+) Ctrl., positive control.