Fig. 6
Acetate treatment mediated IFN-β response in pulmonary cells lines reducing viral load. a–d A549 cells were pre-treated with 260 µM acetate for 24 h and infected with RSV for 96 h. a Percent of PI (propidium iodide) positive cells detected by flow cytometry. b RSV RNA levels detected using real-time PCR (2-ΔCt). c Quantification of RSV PFUs. d Representative images of viral titration assay indicating viral plaques. Lysis plate titration was performed using an anti-RSV antibody. e IFN-β protein levels in supernatants of A549 cells pre-treated with 260 µM acetate for 24 h and infected with RSV for further 24 h. f Cellular viability of A549 WT and A549 Ifnar1-/- assessed using PI by flow cytometry. g RSV RNA levels detected using real-time PCR (2-ΔCt analysis). h Quantification of RSV by PFU assay. i Quantification of translocated NF-kB p65 subunit in A549 cells pre-treated with 260 µM acetate 24 h and infected for 2 h (n = 2 experiments in sextuplicates). j Fluorescence images of NF-kB p65 (green—A) and cell nuclei using DAPI (blue—B). Panel C shows data co-localization (merge). Scale bars = 20 µm. k Western blot analysis of NF-kB p65 in cytoplasmic and nuclear lysates. Cytoplasmic protein bands were normalized by β-actin and nuclear protein bands were normalized by PCNA. L-N, A549 cells were pre-treated 1 h with 2 µM BAY 11–7085. Cells were then treated with 260 µM acetate for 24 h and infected. l Cell viability was evaluated by MTT assay. m RSV RNA levels detected using real-time PCR. n IFN-β protein levels detected by ELISA in the culture supernatant 24 h after infection. All data are expressed as mean ± SEM. Data in a, f, h, m are quadruplicate mean from 3 experiments. Data in b, g, l, m are sextuplicate mean from two experiments. * Significant difference relative to ctrl; # significant difference relative to RSV. Statistical significance was determined with Kruskal–Wallis, except in b, c, g, h, in which Mann–Whitney. *p < 0.05, **p < 0.01, ***p < 0.001. Data in a, b, c, e, f, g, h, i, k, m are provided as a Source Data file
