Figure 2.

Calpain-Cleaved p33 and Recombinant p17 Is More Active than p33 IL-1α

(A) Cytokine concentrations in conditioned media of VSMCs treated with p33 or calpain-cleaved p33, ± IL-1α neutralizing antibody (α pAb).

(B) VSMCs were also treated with calpain sham reactions (no p33) or commercial recombinant p17 IL-1α (cr17) ± calpain sham.

(C) IL-2 concentration in conditioned media of murine EL4 cells treated with p33 ± calpain.

(D) Immunoblot of in vitro cleavage of p33 by calpain.

(E) Coomassie stain of purified soluble p17- and p33-GST fusion proteins.

(F and G) IL-2 (F) or IL-6 (G) concentrations in conditioned media of EL4 or VSMCs, respectively, treated with 1 nM p17- or p33-GST ± α pAb.

(H) IL-2 concentrations in conditioned media of EL4 cells incubated with p33-GST ± calpain, and ± α pAb.

(I) GR1⁺ cells recruited intraperitoneally in wild-type or Il1r1^−/− mice injected with saline or 29 fmol/g p17 or p33.

Data represent mean ± SD or mean ± SEM (I); ^∗p ≤ 0.0003, n = 4 (A), n = 3 (B, C) protein preparations and cleavage reactions; ^∗∗p ≤ 0.007, n = 4. NS, not significant.