Fig 1: The stromal inflammasome controls HSC surface TNFR1/II and MIP receptors via Ras.a, Representative cytokine array blots of WT (left) vs Casp1−/−Casp11−/− (right) bulk bone marrow after 6 h incubation including array controls of kit Membrane C3. b, Concentration of soluble sTNFR1 and sTNFRII, TNF and indicated chemokines measured by ELISA at 6 h post ex vivo culture of bone marrow from adult male WT, Casp1−/−Casp11−/−, and Eμ-myc Casp1−/−Casp11−/− mice or WT bone marrow treated with pan caspase inhibitor QVD-OPH for 2 h. n=3 biological replicates per genotype or condition, ns, p>0.05, p*≤0.05, p**≤0.01, p***≤0.001, p****≤0.0001, 2 WAY Anova. c, Representative histograms of indicated receptor expression on HSC (CD45+Lin–Sca1+c-Kit+CD48–) from (b) at 6 h post ex vivo culture. d, Quantification of gMFI of (c). e, Representative histograms of indicated receptor expression on enriched HSC (CD45+ Lin–Sca1+c-Kit+CD48–) at 16 h of coculture of WT HSC or 4 h Ras inhibitor treated WT HSC with enriched stroma (CD45–Sca-1–c-Kit–Lin–) from WT, Casp1−/−Casp11−/− mice or WT stroma treated with pan caspase inhibitor QVD-OPH for 2 h at 1:2 ratio (HSC:Stroma); n=3 biological replicates per genotype or condition, ns, p>0.05, p****≤0.0001, 2 WAY Anova. f, Quantification of gMFI of (e); n=3 biological replicates per genotype or condition, p*≤0.05, p****≤0.0001, ordinary 2 WAY Anova with Tukey’s multiple comparisons posttest comparing simple genotype column effects within rows. g, Concentration of sTNFR1, sTNFRII, and TNF from (e) cocultures as measured by ELISA; n=3 biological replicates per genotype or condition, ns, p >0.05, p*≤0.05, p**≤0.01, p****≤0.0001, 2 WAY Anova. h, Concentration of indicated MIP family chemokines from (e) cocultures as measured by ELISA; n=3 biological replicates per genotype or condition, ns, p>0.05, p****≤0.0001, 2 WAY Anova. b,d, Both share the same legend shown in b. g,f,h, All share the same legend shown in f. For f, only p values ≤0.05 are shown. All data are presented as mean values +/- SD.
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