(A,B) Spontaneous lifting/licking pain over 1 h after injection of vehicle, S. pyogenes M1 or M3 (5×108 cfu) wt, ΔsagA, Δslo, or ΔsloΔsagA strains (M1, n=8/group; M3, n=12/group). (C,D) Mechanical (n=10/group) and heat (n=9–10/group) sensitivity after injection of S. pyogenes M3 wt or isogenic mutants (5×107 cfu). (E) Spontaneous pain over 1 h in mice injected with S. pyogenes M1 (5×108 cfu) and treated with anti-SLS or control IgG (n=4–5/group). (F) Spontaneous pain over 1 h after injection of S. pyogenes M1 (5×108 cfu) wt or isogenic mutants complemented with plasmid encoding sagA (pDL:sagA) or empty plasmid (pDL278) (n=8/group). Statistical analysis: (A,B,E,F) One-way ANOVA, Tukey post-tests, **p<0.01 ***p<0.001 ****p<0.0001; (C,D) Two-way ANOVA, Bonferroni post-tests, ΔsagA vs wt *p<0.05 ***p<0.001 ****p<0.0001, ΔsloΔsagA vs wt ††††p<0.0001. ns=not significant. nd=not detected. Mean±SEM. See Figure S3 for related data.