Figures
Tables
Supplementary material
Please note: supplementary material is not edited by the Editorial Office, and is uploaded as it has been supplied by the author.
- Supplementary figure S1 - SCFAs modulate iNOS expression. iNOS and GAPDH expression in A549 cells pre-incubated for 1 hour with different concentrations of SCFAs (acetate; 0.5 mM, 2.5 mM, 25 mM, 50 mM) and stimulated with cytomix (CM: IFN-γ, TNF-α, IL-1β; 10 ng/ml each) for A) 4 hours, B) 8 hours, C) 24 hours. Representative Western blots or D) quantification of at least two independent experiments performed in duplicate. Samples and controls for each timepoint were run on the same gel. Fold change relative to cytokine stimulation alone (CM) is displayed.
- Supplementary figure S2 - SCFAs modulate iNOS expression. iNOS and GAPDH expression in A549 cells pre-incubated for 1 hour with different concentrations of SCFAs (propionate; 0.5 mM, 2.5 mM, 25 mM, 50 mM) and stimulated with cytomix (CM: FN-γ, TNF-α, IL-1β; 10 ng/ml each) for A) 4 hours, B) 8 hours, C) 24 hours. Representative Western blots or D) quantification of at least two independent experiments performed in duplicate. Samples and controls for each timepoint were run on the same gel. Fold change relative to cytokine stimulation alone (CM) is displayed.
- Supplementary figure S3 - Effect of acetate concentration and pH on the growth of PAO1. P. aeruginosa PAO1 was incubated with acetate at concentrations between 3.125 mM and 100 mM in TSB. pH was adjusted with HCl to A) 7.0, B) 6.5, C) 6.0, D) 5.5. OD readings at 600 nm were normalised to the average OD from TSB alone at each timepoint. Experiments were performed in duplicate and are representative of at least two independent experiments.
- Supplementary figure S4 - Effect of butyrate concentration and pH on the growth of PAO1. P. aeruginosa PAO1 was incubated with butyrate at concentrations between 3.125 mM and 100 mM in TSB. pH was adjusted with HCl to A) 7.0, B) 6.5, C) 6.0, D) 5.5. OD readings at 600 nm were normalised to the average OD from TSB alone at each timepoint. Experiments were performed in duplicate and are representative of at least two independent experiments.
- Supplementary figure S5 - Effect of propionate concentration and pH on the growth of PA508. P. aeruginosa PA508 was incubated with propionate at concentrations between 3.125 mM and 100 mM in TSB. pH was adjusted with HCl to A) 7.0, B) 6.5, C) 6.0, D) 5.5. OD readings at 600 nm were normalised to the average OD from TSB alone at each timepoint. Experiments were performed in duplicate and are representative of at least two independent experiments.
- Supplementary figure S6 - Effect of propionate concentration and pH on the growth of PAO1 under microaerobic conditions. P. aeruginosa PAO1 was incubated with propionate at concentrations between 3.125 mM and 100 mM in TSB. pH was adjusted with HCl to A) 7.0, B) 6.5, C) 6.0, D) 5.5. Assay plates were sealed with optically-clear PCR film to simulate reduced oxygen environment. OD readings at 600 nm minus 800 nm were normalised to the average OD from TSB alone at each timepoint. Experiments were performed in duplicate and are representative of at least two independent experiments.
- Supplementary figure S7 - High concentrations of SCFAs inhibit A549 cell proliferation but not viability. A) Sub-confluent A549 cells were incubated with SCFAs for 24 hours and measured for proliferation by Prestoblue reagent. Relative fluorescent signal normalised to untreated control is shown. Representative data from one of two experiments. B) Confluent monolayers of A549 cells were incubated with SCFAs for 24 hours and measured for viability by Prestoblue reagent. Mean and SEM of relative fluorescent signal normalised to untreated control is shown. Results of non-parametric ANOVA with Dunn's post-test: *, p<0.05; **, p<0.01.
Disclosures