Journal: Microbiology Spectrum
Article Title: Bacterial Lipoproteins Shift Cellular Metabolism to Glycolysis in Macrophages Causing Bone Erosion
doi: 10.1128/spectrum.04293-22
Figure Lengend Snippet: P2C induces significantly higher cytokine production than P3C in BMDM and MM6 cells. Cytokine production of BMDM (A, B) and BMM (C, D) stimulated with 100 ng/mL of indicated LPP analogues or LPS was assessed as described in Materials and Methods. Control samples (unsti.) were used without adding any stimulators. mTNF-α and mIL6 were measured by ELISA from supernatant of 20 h stimulation. The data were obtained from cells isolated from 3 to 6 mice. Error bars indicate mean ± SEM. Statistical significances were calculated between the treated cells compared to control (non) by one-way ANOVA using Tukey’s multiple-comparison test: *, P < 0.05; **, P < 0.01; ***, P < 0.001. (E to G) Cytokine production of MM6 stimulated with 100 ng/mL of indicated LPP analogues or LPS was assessed as described in Materials and Methods. hTNF-α values were measured by ELISA from supernatant of 5 h stimulation, and hIL6 and hIL-10 were measured by ELISA from supernatant of 18 h stimulation. Three independent experiments were carried out in triplicate. Error bars indicate SEM. Statistical significances were calculated between the treated cells and control (unsti.) by one-way ANOVA using Tukey’s multiple-comparison test; *, P ≤ 0.05; **, P < 0.01; ***, P < 0.001.
Article Snippet: For the blocking assay in MM6 cells, 5 × 10 5 cells were incubated for 1 h with either 1 μg/mL of rabbit anti-human antibodies TNF (catalog no. D1B4; Cell Signaling, Leiden, Netherlands), 1 μg/mL of mouse anti-human antibody IL-6 (catalog no. mabg-hil6-3; InvivoGen, Toulouse, France), 1 μg/mL of rat antihuman antibody IL-10 (catalog no. AHC0103; Thermo Fisher, Schwerte, Germany), or 1 μg/mL mouse anti-human monoclonal antibody IgG1 (catalog no. bgal-mab1-ctrlm; InvivoGen, Toulouse, France) as isotype control.
Techniques: Analogues, Control, Enzyme-linked Immunosorbent Assay, Isolation, Comparison
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