Cell viability Cell viability was determined using alamarBlue (Invitrogen). Briefly, cells were seeded in a 96 well plate at 2×105/ml. After 6 hours of cell adherence, cells were treated in the presence and absence of RBE for 24 hours at 37°C, 5% CO2 in maintenance media. Supernatant was removed and alamarBlue was added to media (20 μg/ml). Fluorescence was detected at excitation:

530/25; emission: 590/35 in ELISA plate reader (Bio-Tek Synergy HT, Winooski, VT). Bacterial quantitation RBE doses of 0, 1, 2, 5 and 10 mg/ml were tested for direct effects on Salmonella viability. Bacteria was added to media at a concentration of 2 × 107 CFU/ml and incubated for 6 hours at 37°C. Bacterial suspension was serially diluted, plated on agar plates and counted after 24 hours incubation. Quantitative PCR for TPCA-1 molecular weight Lactobacillus spp DNA was extracted from fecal pellets of control and rice bran fed mice before and SAHA after Salmonella challenge using a MoBio Powersoil DNA extraction kit (MoBio, Carlsbad, CA). A dilution of DNA from pure cultures of Lactobacillus rhamnosus was used to generate standard curves and DNA from Pseudomonas aeruginosa were run as a negative control to ensure primer specificity. DNA was quantified by Nanodrop (Thermo Fisher Scientific) and diluted to 5 ng/μl. Real time PCR primers were used from Malinen et al. [47] for amplification of Lactobacillus spp. Samples were run on an ABI Prism 310 thermocycler (Applied Biosystems)

using the following program: 95°C for 3 min 30 s followed by 30 cycles of 95°C for 15 s, 58°C for 20 s 72°C for 30 s and melt curves MLN4924 were generated by 95°C for 1 min followed by eighty 10 s repeats at set point temperatures incrementally decreasing by 0.5°C. Statistical analysis Data was analyzed using Graphpad Prism5 Software. Experiments

were repeated a minimum of three times. GNA12 Raw data were log transformed into a log10 scale for CFU analysis and repeated measures ANOVA and post hoc Tukey’s test were used for Salmonella fecal shedding and fecal Lactobacilli measures. Inflammatory cytokines were analyzed using two -way ANOVA and Bonferroni post hoc test. A nonparametric ANOVA (Kruskal Wallis) was performed, followed by Dunn’s test for in vitro Salmonella assays. Significance was determined for all studies at P <0.05. Acknowledgements We would like to thank Dr. Andres Vazquez-Torres for providing the strain of Salmonella used in these studies, and Dr. Anna McClung from the USDA-ARS Dale Bumpers Rice Research Center for providing rice bran from the single Neptune variety. We also thank Dr. Daniel Manter from USDA-ARS-Soil Plant Nutrient Research, Brittany Barnett for for assistance with qPCR of Lactobacillus spp. and Adam Heuberger and Caleb Schmidt for their technical assistance. Funding A Grand Explorations in Global Health Grant from the Bill and Melinda Gates Foundation (OPP1015267) and the Shipley Foundation supported this work.