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. 2019 Nov 26;13(2):322–333. doi: 10.1038/s41385-019-0235-4

Fig. 5. SerpinA3N is secreted during infection and inhibits NE activity.

Fig. 5

a Western blot analysis of faecal samples taken from three uninfected and three infected C57 (8 dpi) and C3H (6 dpi) mice using anti-SerpinA3N antibody. Free SerpinA3N (ca. 40 kDa) is detected in uninfected mice and infected C57. In both C57 and C3H mice, a band at ~55 kDa (arrow) corresponds to a protease–SerpinA3N complex. Bar graph below blot is quantification band/s intensity; actual values are indicated in italics above each bar. b Faecal samples from infected C57 mice (1 mg/ml- triangles) significantly inhibit recombinant NE (rNE) activity whereas BSA (circles) or lower concentrations (0.1mg/ml-inverted triangles; 0.01mg/ml-diamonds) did not. Recombinant SerpinA3N inhibits rNE (squares). Grey circles are NE activity in faecal samples from C57 mice without rNE. Multiple comparison one-way ANOVA, n.s. = non-significant, ***P ≤ 0.001. c Rate of NE substrate cleavage by rNE pre-incubated with faecal samples from C3H (squares) or BSA (circles). Multiple comparison one-way ANOVA, n.s. = non-significant, **P ≤ 0.01, ***P ≤ 0.001. d Western blot analysis of recombinant (r) SerpinA3N incubated with or without activated rNE. Band at ca. 75 kDa is complexed SerpinA3N and NE, which is present in both SerpinA3N and NE blots.