Supplemental Data
A role for the gut microbiota and the metabolite sensing receptor GPR43 in the pathogenesis of acute Gout
Angélica T. Vieira1,2,3, Laurence Macia1, Izabela Galvão2, Flaviano S. Martins4, Maria Cecília C. Canesso1,2, Flávio A. Amaral2, Cristiana C. Garcia2, Ellen De Leon3, Doris Shim3, Kendle Maslowski3, Jacques Nicoli4, Leda Q. Vieira2, Jacquie L. Harper5, Mauro M. Teixeira2* and Charles R. Mackay1*
1School of Biological Sciences, Faculty of Medicine, Nursing and Health Sciences, Monash University, Wellington Road, Clayton, Victoria 3800, Australia; 2Immunopharmacology Group-Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais, Brazil; 3Garvan Institute of Medical Research, 384 Victoria Street, Darlinghurst, New South Wales 2010, Australia 4Department of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais, Brazil 5Malaghan Institute of Medical Research, Wellington 6242, New Zealand.
Supplemental data- 1
Supplemental data-1. Intestinal microbiota depletion using antibiotic therapy in conventional showed the same hyporresponsiveness observed in GF mice after gout induction. Conventional mice treated or not with were injected with MSU crystals (100μg/ knee joint) and number of (A) total cells and (B) number of neutrophils 15hr after injection were assessed in the articular cavity. A combination of large spectrum antibiotics was used to eliminate the intestinal microbiota of conventional mice. To this end a combination of ampicillin 2 g/L, neomycin 2 g/L, metronidazole 1 g/L (and vancomycin 0.5 g/L was administrated in the drinking water and concomitantly, all mice received an intra-peritoneal injection of ciprofloxacin (0.2 g/L) during 21 days prior to MSU injection. Results are expressed as mean ± SEM of 5-7 mice per group
Supplemental data- 2
Supplemental data- 2. ROS production by macrophages after LPS exposition was not different between Germ free, Gpr43-/- and conventional.. Reactive Oxygen Species -ROS production was measured in non-primed peritoneal macrophages isolated from WT (Conventional wild type) and Gpr43-/- (KO) and GF mice. Macrophages was incubated with Lipopolysaccharide LPS (μg/ml) and measured during the time as indicated in the graphic. These results are expressed as mean ± SEM of 5-7 mice per group.
