Effect of extracellular potassium on IL-1 processing and release.

<p>A) Bone marrow-derived macrophages were plated in triplicate wells in 12-well plates and primed with 50 ng/ml of LPS for 4 h. Cells were then incubated in medium containing 130 mM NaCl/5 mM KCl or 5 mM KCl/130 mM NaCl in the presence of absence of 0.01 µg/mL ricin, 25 µg/mL cycloheximide, 10 µg/mL, 10 µg/mL emetine, 75 µg/mL puromycin, 0.2 µg/mL pactamycin, 10 µg/mL anisomycin, 3.4 µM nigericin, or 5 mM ATP for 4 h. Medium was collected, and p17 IL-1 was determined by ELISA. B) Macrophages were plated, primed with LPS and incubated in medium containing 130 mM NaCl/5 mM KCl (MEM-Na) or 5 mM NaKCl/130 mM KCl (MEM-K) in the presence or absence 5 mM ATP or 3.4 µM nigericin for 4 h. Proteins were precipitated from the media with TCA and analyzed by Western blotting. C) Macrophages were plated, primed with LPS, and incubated in medium containing 130 mM NaCl/5 mM KCl (MEM-Na) or 5 mM NaKCl/130 mM KCl (MEM-K) in the presence or absence 10 ng/mL ricin, 10 µg/mL emetine, or 25 µg/mL cycloheximide for 4 h. Proteins were precipitated with TCA and analyzed by Western blotting.