Measurements were conducted in duplicates and the fold
increase expression MLN2238 cell line was calculated by using the expression 2^DCt, according to the instructions from Applied Biosystems User’s Bulletin #2 (P/N 4303859). Results were shown as mean values ± standard deviation. Statistical analysis The means of the groups were evaluated by analysis of variance (ANOVA) followed by the Dunn’s or Bonferroni’s post test. A probability value of less than 0.05 was considered statistically significant, and all the comparisons were performed using the GraphPad Prism 5.00 software (GraphPad Software, San Diego California, USA). Acknowledgments A.D.P. was supported by a fellowship from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Grant Number AUXPE/PNPD 2439/2011, Brasília, Brazil). This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico BI 6727 (Grant
Number 201179/2009-1) and by a Grant from the Fundação de Amparo à Pesquisa de Minas Gerais (Belo Horizonte, Brazil). Electronic supplementary material Additional file 1: Cytokine production in spleen of five-week old female BALB/c mice treated with bovicin HC5 or ovalbumin. The relative expression of IL-12p40 (A), IFN-γ (B), IL-5 (C), IL-13 (D), TNF-α (E), TGF-β (F), IL-10 (G), IL-4 (H), IL-17 (I) mRNA was determined by real time-PCR and calculated by reference to the β-actin in each sample, using the threshold cycle (Ct) method. Results are shown as the mean value ± SD of duplicate samples from three independent mice within the NC, Bov and PC groups. Differences among treatments were indicated by different lowercase letters and were considered statistically significant by the Bonferroni multiple comparison
test (p < 0.05). (NC) negative control group; (Bov) mice treated with bovicin HC5; (PC) positive control group. (TIFF 10328 kb) (TIFF 10 MB) References 1. Delves-Broughton J: Nisin as a food preservative. Food Aust 2005, 57:525–527. 2. Gálvez A, López RL, Abriouel H, Valdivia E, Ben ON: Application of bacteriocins in the control of foodborne pathogenic and Lepirudin spoilage bacteria. Crit Rev Biotechnol 2008, 28:125–152.PubMedCrossRef 3. Gänzle MG, Weber S, Hammes WP: Effect of ecological factors on the inhibitory spectrum and activity of bacteriocins. Int J Food Microbiol 1999, 46:207–217.PubMedCrossRef 4. Toke O: Antimicrobial peptides: new candidates in the fight against bacterial infections. Biopolymers 2005, 580:717–735.CrossRef 5. Belguesmia Y, Madi A, Sperandio D, Merieau A, Feuilloley M, Prévost H, Drider D, Connil N: Growing insights into the safety of bacteriocins: the case of enterocin S37. Res Microbiol 2011, 162:159–163.PubMedCrossRef 6. Pariza MW, Foster EM: Determining the safety of enzyme used in food processing. J Food Protect 1983, 46:453–468. 7. Pariza MW, Cook M: Determining the safety of enzymes used in animal feed. Regul Toxicol Pharmacol 2010, 56:332–342.PubMedCrossRef 8. FDA. U.S.