Figure 3: Obesity-induced NLRC4 inflammasome in tumour-infiltrating myeloid cells.

(a) Data represent the number of tumour-infiltrating total (mφ), CD11c⁻ or CD11c⁺ macrophages as a percentage of total cells counted±s.d. (n=5 ND; n=9 HFD). (b) mRNA expression (relative to peptidylprolyl isomerase A gene; Ppia) of the indicated genes in tumours from indicated mice (n=3)±s.d. (c) Western blot analysis for CASP1 (left panels) and CASP11 (right panels) in tumour-infiltrating CD11b⁺ and CD11b⁻ cell populations. Cells were combined from four to five tumours from each group. (d) mRNA fold-change relative to the ND group, using Ppia as reference gene. Mean±s.d. in tumours from the indicated mice (n=3). (e,f) Nlrc4 mRNA expression in tumour-infiltrating CD11b⁺ and CD11b⁻ cell populations from the indicated mice in the indicated tumour model, using Actin beta (Actb) as the reference gene. Cells were combined from four to five tumours from each group, and data are shown in triplicates. (g) Western blot for NLRC4-flag in tumour-infiltrating CD11b⁻ and CD11b⁺ cell populations from DIO mice. (h) Data represent the average number of tumour-infiltrating CD45⁺ cells with CASP1 activation as a percentage of CD45⁺ cells±s.d. (n=5 WT ND, n=5 WT HFD, n=5 Nlrc4^−/− ND, n=4 Nlrc4^−/− HFD). For all panels, Student’s t-test was used to determine significance. (i) NLRC4 activation from macrophages drives ODBP. Bone marrow macrophages from WT or Nlrc4^−/− female mice were co-injected with Py8119 cells orthotopically into DIO Nlrc4^−/− female mice. Tumour growth was monitored weekly (n=7, means±s.e.m.). Tumour study was repeated in a different cohort of animals. All other studies represent results from two to three repeats.