Figure 1.

B. dentium γ-glutamylcysteine enter host cells, upregulate glutathione and reduce ROS, NF-kB, and cytokine synthesis. a. Fluorescein-5-Maleimide was used to fluorescently tag cysteine residues in y-glutamylcysteine, B. dentium conditioned LDM4 media, or uninoculated LDM4 media. Representative histograms from flow cytometry analysis of T84 cells after exposure to cysteine-tagged y-glutamylcysteine, B. dentium-conditioned LDM4 media, or uninoculated LDM4 media (control) (n = 3/experiment). b. Representative images of T84 cells following incubation with Fluorescein-5-Maleimide-tagged B. dentium conditioned LDM4 (which fluorescently labels cysteine residues), counterstained with nuclear dye Hoechst (scale bar = 50 µm). c. Measurement of glutathione levels in T84 cells after 3 hr using a Thiol-tracker, as measured on a fluorescence plate reader (ex/em: 405/528) (n = 3/experiment). d. Measurement of ROS levels in T84 cells after 3 hr in cells stained with H₂DCFDA, as measured on a fluorescent plate reader (ex/em: 485/528) (n = 3/experiment). e. Secreted NF-kB luciferase quantified from T84 monolayers treated for 16 hr (n = 4/experiment). f. IL-8 levels of T84 cells after 16 hr incubation with treatment as measured by ELISA(n = 3/experiment). All data is expressed as mean ± st dev and all experiments were repeated 3–4 independent times. *p < .05, Multi-Way ANOVA