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. Author manuscript; available in PMC: 2018 Mar 22.
Published in final edited form as: Cell. 2016 Sep 22;167(2):457–470.e13. doi: 10.1016/j.cell.2016.08.064

Figure 2. Inhibition of succinate dehydrogenase impairs LPS-induced IL-1β production and boosts IL-1RA and IL-10.

Figure 2

Figure 2

BMDMs were pretreated for 3 h with dimethyl malonate (DMM; 10 mM) prior to stimulation with LPS (100ng/ml) for 24 h (A), 48 h (B, C, E, G, H), or 4 h (D, F). SDHB-proficient (SDHBfl/fl EtOH) and SDHB-deficient (SDHBfl/fl Tamox) BMDMs were untreated (Ctl) or treated with LPS (100 ng/μl) for 24 h (I, J). Lysed cells were analyzed by liquid chromatography-mass spectrometry (LC-MS) to determine succinate levels (A). mRNA from total cell lysates was analyzed by qPCR for IL-1β (B), TNF-α (D), IL-1RA (F) and IL-10 (G) expression. Whole cell lysates were analyzed by Western blotting for pro-IL-1β, HIF-1α and β-actin (C, I). Supernatants were analyzed by ELISA for TNF-α (E, J) and IL-10 production (H). The data in (A, B, D - H) represent mean ± S.E.M., n=3. The data in (J) represent mean ± S.E.M., n=6, *p < 0.05, **p < 0.01, ***p < 0.001. The blots are representative of 3 (C) or 5 (J) independent experiments.