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. Author manuscript; available in PMC: 2023 Apr 29.
Published in final edited form as: Circ Res. 2022 Apr 11;130(9):1289–1305. doi: 10.1161/CIRCRESAHA.121.320704

Figure 4. PKM2 deficient macrophages exhibit reduced pro-inflammatory response and reduced glycolytic activity.

Figure 4.

All the mice were females. A, Real-time quantitative PCR analysis of pro-inflammatory and anti-inflammatory genes in primary elicited macrophages of mice fed a high-fat “Western” diet (n=7, 7). B, Bone marrow-derived macrophages (BMDMs) from wild-type Ldlr−/− mice were unstimulated (n=7) or stimulated with 100 ng/mL LPS (n=7) or 10 ng/mL IL-4 (n=7) for 24 hrs and nuclear and cytosolic PKM2 protein levels were analyzed by immunoblotting. Loading control: Lamin-B1 and GAPDH. C, BMDMs were stimulated with LPS for 4 hours. Representative immunoblots and densitometric analysis of phospho and total STAT3 in nuclear proteins fraction. Loading control: Lamin-B1. Unstimulated (n=7, 7). LPS-treated (n=7, 7). D, Glycolytic proton efflux rate (glycoPER) levels in untreated and LPS-treated (100 ng/ml) BMDMs (100,000 cells per well) were assessed by Seahorse extracellular flux analyzer. The basal respiration values were noted for 15 mins before injecting the Rot/AA (0.5 μM). Finally, 2-DG was injected, and values were measured for 40 mins. Left panel shows glycoPER curve, whereas right panel shows basal and compensatory glycolysis rate. Unstimulated, (n=10, 10). LPS-treated (n=10, 10). Results were presented with mean ± SEM. Statistical analysis as indicated in the figure panels.