Figure legends
Figure 1. Both porcine pancreatic elastase (PPE) and papain
induced pulmonary emphysema, and papain reproduced the asthmatic
features. (A) Protocol (B) Respiratory mechanics (C) Lung tissue
stained with haematoxylin and eosin (HE; 20x) or (E) Alcian blue and
periodic acid-Schiff (AB–PAS; 20x) (D) Calculation of the mean linear
intercept (F) Representative bronchoalveolar lavage fluid (BALF)
cytospin preparations (50x); M, macrophage; E, eosinophil. (G)
Differential cell counts of cytospin preparations (H) Airway
responsiveness to increasing doses of methacholine measured by
subtracting the baseline from each airway resistance; *p< 0.05, **p < 0.01, ***p <
0.001; n.s. not significant.
Figure 2. Papain induced an increase in the
macrophage/neutrophil- and type 2 inflammation-associated cytokines;
allergic airway inflammation and emphysema persisted for 5 weeks after
the final papain administration. (A) Cytokines in the BALF (B) Total
serum IgE (C) Neutrophil gelatinase-associated lipocalin (NGAL) in the
serum and BALF (D) Protocol (E) Differential cell counts from BALF
cytospins (F) Respiratory mechanics (G) Lung tissue stained with HE;
20x. (H, I) Total and papain-specific IgE and NGAL; *p< 0.05, **p < 0.01, ***p <
0.001; ND, not detected; n.s. not significant.
Figure S1. Body weights of mice in the PPE- and papain-treated
groups were not significantly different from those treated with PBS
control. Weights of mice treated with PPE (n = 19), papain (n = 27) or
PBS (n = 20) measured on days 0, 7, 14, and 21 as in Fig. 1A; **p< 0.01; n.s. not significant with two-way repeated measures
ANOVA with Holm-Bonferroni correction.
Figure S2. Papain-induced mucus hypersecretion and allergic
airway inflammation associated with elevated levels of transcript for
Muc5ac, Muc5b, Eotaxin1 and Eotaxin2. Quantitative RT-PCR determination
of whole lung mRNA encoding Muc5ac, Muc5b, Eotaxin1 and Eotaxin2;
*p < 0.05, **p < 0.01, ***p< 0.001; n.s. not significant.
Figure S3. IL-33 in serum and BALF on day 25. Concentrations of
IL-33 in serum and BALF measured by ELISA; n.s. not significant.
Figure S4. Administration of poly(I:C) augmented airway
inflammation and NGAL in the PPE- and papain-treated mice; allergic
airway inflammation was sustained in the papain-treated group. (A)
Protocol (B) Representative BALF cytospins; L, lymphocyte; N,
neutrophil; E, eosinophil (50x) (C) Differential cell counts from BALF
cytospins (D) Lung tissue stained with HE (20x) (E) Cytokines in BALF
(F) NGAL in the serum and BALF; *p < 0.05, **p< 0.01, ***p < 0.001; n.s. not significant;
ND, not detected.
Table S1. Cytokine array data. GM-CSF, Granulocyte macrophage
colony-stimulating factor; IFN-γ, Interferon gamma; IL, Interleukin; KC,
Keratinocyte-derived chemokine (CXCL1); MCP-1, Monocyte chemotactic
protein-1 (CCL2); M-CSF, Macrophage colony-stimulating factor; RANTES,
Regulated on activation, normal T cell expressed and secreted (CCL5);
TNF-α, Tumour necrosis factor alpha; VEGF, Vascular endothelial growth
factor. The data are presented as the mean (±SEM). n = 3 per group; ND,
not detected.