Figure 3. Microbiota-mediated regulation of IL-1β expression shapes inflammatory bone disease.

a, qRT-PCR analysis of relative proIl1β expression in the footpads of 12-16 week old WT, LFD Pstpip2^cmo and HFD Pstpip2^cmo mice. Each point represents an individual mouse, and the line represents the mean ± s.e.m. Combined data from three independent experiments. b, Protein levels of IL-1β in the hind paws. Combined data from two independent experiments. c, Relative proIl1β mRNA expression levels in CD45⁺ cells isolated from the colons of specific pathogen free (SPF) and germ-free (GF) WT mice. Two biological replicates, with two technical replicates each. d-e, Pstpip2^cmo mice were treated with a cocktail of broad-spectrum antibiotics in their drinking water (Abx). d, qRT-PCR analysis of colonic Il1β expression levels from 12-14 week old Pstpip2^cmo mice that received either regular drinking water (n=15) or Abx water (n=9). e, Incidence of inflammatory bone disease. f-i, Young Pstpip2^cmo mice (3 wks old) received PBS or fecal microbiota from diseased LFD Pstpip2^cmo or disease-free HFD Pstpip2^cmo mice by oral transplantation. f, 16S rDNA analysis of Prevotella copy numbers. g, Incidence of inflammatory bone disease. h-i, Representative footpad images (h) and H&E micrographs (i). (*P < 0.05, **P < 0.01, ***P < 0.001; Student’s t-test)