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. Author manuscript; available in PMC: 2017 Oct 1.
Published in final edited form as: J Immunol. 2016 Aug 24;197(7):2909–2917. doi: 10.4049/jimmunol.1600651

FIGURE 2. LA effects are pH- and MCT-1-dependent.

FIGURE 2

A) BMMC were cultured in media alone, 12.5 mM lactic acid, or sodium lactate media for 24 hours prior to IL-33 activation. ELISA was used to measure IL-6, TNF, IL-13, and MCP-1 in culture supernatants. B) BMMC were treated with either vehicle (DMSO) or the indicated MCT inhibitors, α-cyano-4-hydroxycinnamic acid (CHC) or AR-C155858 for 1 hour prior to lactic acid or media treatment for 24 hours. Cells were then activated with 100ng/mL of IL-33 for 16 hours, supernatants collected, and IL-6 measured using ELISA. C) MCT-1 and MCT-2 expression was measured during 40 cycles of RT-qPCR using RNA harvested from BMMC. β-actin was used as the housekeeping gene for MCT-1 and GAPDH for MCT-2, based on primer optimization and melting point similarities. ND means “not detectable”. Results are expressed as mean ±SEM and are representative of 3 independent experiments conducted in triplicate. *, p<0.05; **, p<0.01; ***, p<0.001; ****, p<0.0001; NS, not significant.