Fig. S1. Representative images of composites from the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay. Alveolar macrophages from three chronic obstructive pulmonary disease (COPD) patients were treated with and without cigarette smoke extract (CSE) [0·1/0·2 optical density (OD)] for 2 h followed by an additional 24 h with CSE-free medium; 1% Triton X was used as a positive cell death control and the 0·2 OD CSE condition was used for the negative TUNEL control [0·2 OD (−)]. Blue fluorescence illustrates 4′,6-diamidino-2-phenylindole (DAPI) labelling of DNA in the nucleus and green fluorescence illustrates TUNEL labelling of DNA fragmentation.


Fig. S2. Dose-dependent time–course of Toll-like receptor (TLR) ligand-induced interleukin 8 (CXCL8)/tumour necrosis factor (TNF)-α secretion from chronic obstructive pulmonary disease (COPD) alveolar macrophages. Alveolar macrophages from six COPD patients (two current and seven ex-smokers) were treated with varying doses of phase I flagellin (FliC) (a,d), ultra-pure Escherichia coli lipopolysaccharide (UPLPS) (b,e) and Pam3CSK4 (PAM) (c,f) during 48 h. Collected supernatants were analysed for CXCL8 (a–c) and TNF-α (d–f) by enzyme-linked immunosorbent assay (ELISA). Data sets from each time-point were analysed using the Friedman test and Dunn's multiple comparison test. *, **, *** = significant difference compared to matched medium control (P < 0·05, P < 0·01 and P < 0·001, respectively).

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