Figure 5. TRIM21 is involved in inflammasome activation rather than enhancing pro‐IL‐1β expression.

1. HMDM were stimulated with AdV (50,000 pp/cell) and antibody (h9C12; 20 μg/ml, IVIg: 20 mg/ml) or 10 μg/ml pI:C for 3 h and gene expression was measured by qPCR (n = 4 mean ± s.e.m).
2. HMDM were stimulated as in (A), or with 10 ng/ml LPS for 6 h and pro‐IL‐1β levels in the cytosol measured by Western blot. Blot is representative of three independent donors.
3. HMDM were stimulated as in (A), or with 10 ng/ml LPS for 3 h and NLRP3 mRNA expression measured by qPCR. Data show average ± s.d. of two independent donors.
4. HMDM were stimulated as in (B) and NLRP3 levels in the cytosol measured by Western blot. Blot is representative of two independent donors.
5. TLR9 mRNA levels from PBMCs, CD19^+ve B cells, CD14^+ve monocytes or HMDM derived from CD14^+ve monocytes were assessed by qPCR, and copy number was determined relative to actin copy number (n = 5 mean ± s.e.m).
6. THP‐1s expressing ASC‐GFP were stimulated for 6 h with virus and antibody or 200 ng/well HT‐DNA or 10 μM Nigericin, in the presence of the pan‐caspase inhibitor zVAD‐fmk. A representative image (scale bar 100 μm) and quantification of number of cells with ASC specks from three independent experiments (mean ± s.e.m, *P ≤ 0.05, paired, two‐tailed t‐test) are shown.

Source data are available online for this figure.