Figure S6.

Autoactivation of BMDMs and the Effect of Antibiotics on Systemic Inflammation in OTULIN-Deficient Bone Marrow Chimeric Mice, Related to Figure 6

(A) Immunoblot showing NF-κB activation in MEFs treated with 0.25 ng/mL TNF and anti-TNF neutralizing antibodies or isotype control as indicated. (B) Schematic of the experiment indicating timing of enrofloxacin and tamoxifen treatment. (C) Enrofloxacin treatment does not rescue weight loss, indicating sterile inflammation. Tamoxifen (tx) was administered i.p. (arrows) to CreERT2-Otulin^flox chimeras treated with enrofloxacin or not. (D) Blood cell counts taken at day 4 from CreERT2-Otulin^flox chimeras treated with enrofloxacin or not showing that enrofloxacin does not reduce neutrophilia in CreERT2-Otulin^LacZ/flox chimeras, indicating sterile inflammation. (E) Luminex multiplex analysis of cytokine and chemokine concentrations in serum from terminal bleeds on day 4 of CreERT2-Otulin^flox chimeras treated with enrofloxacin or not showing that enrofloxacin does not reduce secretion of inflammatory cytokines in CreERT2-Otulin^LacZ/flox chimeras, indicating sterile inflammation. (B-E) Data were pooled from two independent experiments (except for G-CSF concentrations in E). (F) Micrographs of H&E stained sections of livers reveal inflammatory foci (arrowheads) in liver parenchyma showing that enrofloxacin does not reduce infiltration and inflammatory foci in CreERT2-Otulin^LacZ/flox chimeras, indicating sterile inflammation. Micrographs are representative of 10 CreERT2-Otulin^+/flox and 12 CreERT2-Otulin^LacZ/flox chimeras from two independent experiments. Scale bars: 200 μm. (G) Frequency of infiltrating CD11b⁺Gr-1⁺ neutrophils. (C-E and G) Data are presented as mean ± SEM, and n represents number of mice.