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. 2021 Jun 10;54(7):1463–1477.e11. doi: 10.1016/j.immuni.2021.05.004

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Metformin attenuates Il1b and Il6 mRNA induction but does not inhibit NF-κB activation

(A) BMDM pretreated -/+ metformin (0.5 mM, 16 h) were harvested 30 min after LPS (100 ng/mL) addition. Nuclear (Nuc) and cytosolic (Cyt) fractions were analyzed for the indicated proteins. s.e., short exposure; l.e., long exposure. One representative immunoblot (IB) out of 2.

(B) IB analysis of p62, DRP1, and VDAC in mitochondria from BMDM pretreated -/+ metformin, primed with LPS, and challenged with ATP (4 mM, 1 h).

(C) Q-PCR quantitation of Il1b, Il6 and Il10 mRNAs before or after LPS stimulation -/+ metformin pre-treatment (n = 3).

(D) p65, C/EBPβ or NFATc1-4 recruitments to the Il1 and Il6 promoters in BMDM before or after LPS stimulation was analyzed by ChIP-qPCR assay (n = 3–7).

(E) IB analysis of p38 MAPK and JNK phosphorylation (left) and quantitation of Il6 mRNA (top) and IL-6 secretion (bottom) from BMDM pretreated with metformin, SB202190 (10 μM) or SP600125 (40 μM) for 16 h and stimulated with LPS for 30 min.

All IB show one representative out of 3. Results in (C–E) are averages ±SD. ^∗p < 0.05; ^∗∗p < 0.01; ^∗∗∗p < 0.001. Two-sided unpaired t test. See also Figure S1.

Figure S1