Inhibition of Extracellular Calcium Influx Results in Enhanced IL-12 Production in LPS-Treated Murine Macrophages by Downregulation of the CaMKK-AMPK-SIRT1 Signaling Pathway

Figure 2

Depletion of extracellular calcium selectively upregulates IL-12 expression in LPS-treated murine macrophages. (a, b) Murine peritoneal macrophages cultured in DMEM with or without calcium for 1 h and then stimulated with LPS (100 ng/mL). Supernatants were collected after 24 h and the protein levels of TNF-α, IL-6, and IL-12 p40 were detected by ELISA ((a) ). Total RNA was extracted after 12 h and mRNA expressions of TNF-α, IL-6, and IL-12 p40 were detected by real-time PCR ((b) ). versus LPS. (c, d) Murine peritoneal macrophages were incubated with LPS (100 ng/mL) in normal DMEM, calcium-free DMEM, calcium-free DMEM with 2 mM CaCl₂, or normal DMEM with 5 mM EGTA. The mRNA expressions of IL-12 p40 and IL-12 p35 were detected via real-time PCR (c). The supernatant levels of IL-12 p40 and IL-12 p35 were detected via ELISA (d). versus medium; and versus LPS ().

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