Fig. 7.
Budesonide inhibits the LC-promoting effect of HDM in KrasG12D mice and HDM-induced IL-1β production by macrophages. AKras.G12D mice were treated i.n with VEH and dimethyl sulfoxide (DMSO), HDM and DMSO, or HDM and budesonide (Bud) as indicated in this schematic overview of the study design. VEH + DMSO (n = 4), HDM + DMSO (n = 7), and HDM + Bud (n = 11). B Representative pictures of H&E-stained lung sections of mice treated as in A. Scale bars, 2 mm. C Tumor multiplicity calculated on H&E-stained sections as shown in B. D BMDMs were isolated from WT mice as shown in Supplemental Fig. 4A and were treated for 6 h with VEH (PBS) or HDM (200 μg/mL) and DMSO (0.01%) or with HDM (200 μg/mL) and budesonide (Bud, 1 nM), and the relative mRNA level of Il1b was measured by qPCR and normalized to Gapdh gene expression. The relative mRNA level in the VEH condition was used as a reference and assigned to 1. E WT BMDMs were treated as in D but for 24 h. ATP (5 mM) was added to each well for the last hour of culture and total cell lysates were prepared for western blot analysis of NLRP3 and pro-IL-1β expression. Densitometric measurements of NLRP3 and pro-IL-1β relative to β-actin are indicated below each blot. F The culture supernatants from the cells stimulated in E were recovered and the levels of IL-1β were analyzed by ELISA. Data are representative of two (A–C) or three (D–F) independent experiments and are expressed as mean ± SEM. Statistical significance was assessed by one-way ANOVA with post hoc Bonferroni’s test. n.s: non-significant, * P < 0.05, ** P < 0.01, **** P < 0.0001