Fig. S1. Immunophenotype of induced sputum cells. Single-cell suspensions were prepared from sputum samples and stained with anti-CD45, anti-CD16 and anti-CD3 or anti-CD14. Vital dye 7-aminoactinomycin D (7-AAD) was used to exclude dead cells. Representative flow histogram of an asthmatic patient is shown. Numbers inside dot-plots indicate the percentage of each subpopulation.


Fig. S2. Effect of galectins (gal) on cytokine expression. (a–c). Effect of gal-3 on lipopolysaccharide (LPS)-induced interleukin (IL)-12A (a) and of gal-9 on LPS-induced IL-12B and TNF-α (b,c) expression on peripheral blood mononuclear cells (PBMC) from healthy subjects. PBMC (5 × 105) were incubated on p24 plates with 100 ng/μl LPS in the presence or not of gal-3 or gal-9 (10 μg/ml). After 24 h culture, cytokine expression was analysed by reverse transcription–polymerase chain reaction (RT–PCR). Data correspond to five independent experiments. Difference between treatments was analysed by Wilcoxon's matched-pairs test. (d) Effect of gal-1 and gal-9 on LPS-induced IL-10 expression on peripheral blood mononuclear cells (PBMC). Cells were treated and analysed as in (a–c). (e) Gal-1 and gal-9 induce the expression of IL-10 in PBMC. Mononuclear cells (5 × 105) were incubated on p24 plates in the presence of 10 μg/ml gal-1, gal-3 and gal-9 during 24 h, and then IL-10 expression was determined by RT–PCR. LPS (100 ng/ml) was used as positive control. Data correspond to mean ± standard error of the mean of five independent experiments. Differences among treatment were tested by one-way analysis of variance test, *P < 0·05.


Table S1. Sequence of primers used for reverse transcription–polymerase chain reaction (RT–PCR).

Table S2. Relation between beclomethasone (BDP) dose and levels of protein expression [mean fluorescence intensity (MFI)] by flow cytometry.

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