Skip to main content
. Author manuscript; available in PMC: 2016 Dec 29.
Published in final edited form as: Cell Metab. 2015 Jan 6;21(1):65–80. doi: 10.1016/j.cmet.2014.12.005

Figure 1. Tetramerization of LPS-induced PKM2 in primary BMDMs inhibits the Hif-1α targets IL-1β and Ldha.

Figure 1

LPS-stimulated BMDMs were assayed for expression of PKM2, Y105 phosphorylated PKM2 and β-actin by Western Blotting (A) and Pkm2 mRNA by qRT-PCR (C). (B) Crosslinking (500μM DSS) and western blot of endogenous PKM2 in BMDMs ±LPS (24 hrs). (D) LPS did not significantly affect expression of PKM1 in BMDMs. (E) BMDMs pretreated with ± DASA-58 or ±TEPP-46 as indicated, followed by LPS for 24 hours. Cytosolic and nuclear fractions were isolated and PKM2, β-actin, Lamin A, and Tubulin were detected by western blotting. (F) DSS crosslinking and western blotting of PKM2 in BMDMs and RAW macrophages after treatment ±100μM TEPP-58 or ±50μM DASA-46. (G) RAW macrophages treated with ±10μM TEPP-58 or DMSO (1h), followed by LPS. Protein separated by size exclusion chromatography and blotted for PKM2. (H) BMDMs (left) or PECs (right) were pretreated ±DASA-58 or TEPP-46 (30 min), followed by stimulation with LPS for 24 hours. Cell lysates were analyzed for pro-IL-1β or β-actin expression by western blotting. (I) Il1b (left panel), Il6 (middle panel) and Ldha (right panel) mRNA and (J) IL-6 (left) and TNF (right) protein expression were measured in BMDMs treated with ±DASA-58 and LPS for 6–24 hours. Data represents Mean ± SEM, n=3, **p<0.01.