Skip to Main Content
Table II.

Effect of ethacrynic acid and ATP on IL-1 recoverya

ConditionIL-1βIL-1α
35I28I35e28e17eTotal31I31e17eTotal
Pulse 6075 151 6226 3774 3774 
Chase 3812 82 3894 2871 2871 
Chase + ethacrynic acid 3311 95 3406 2560 2560 
Chase + ATP 759 100 249 180 5851 7139 1196 1902 134 3232 
Chase + ATP+ ethacrynic acid 954 116 92 20 953 2135 1713 338 2051 
ConditionIL-1βIL-1α
35I28I35e28e17eTotal31I31e17eTotal
Pulse 6075 151 6226 3774 3774 
Chase 3812 82 3894 2871 2871 
Chase + ethacrynic acid 3311 95 3406 2560 2560 
Chase + ATP 759 100 249 180 5851 7139 1196 1902 134 3232 
Chase + ATP+ ethacrynic acid 954 116 92 20 953 2135 1713 338 2051 
a

LPS-activated [35S]methionine-labeled macrophages were harvested immediately (Pulse) or treated with 2 mM ATP (where indicated) for 15 min and chased for 3 h in the absence or presence of 10 μM ethacrynic acid. Cell and media samples were harvested separately, each was divided into two equal fractions, and IL-1α and IL-1β were recovered by immunoprecipitation. The resulting immunoprecipitates were separated by SDS-PAGE, and radioactivity associated with individual species was determined. The quantity of radioactivity recovered intracellularly in association with the 35-kDa (35I) and 28-kDa (28I) forms and extracellularly in association with the 35-kDa (35e), 28-kDa (28e), and 17-kDa (17e) forms of IL-1β are indicated; counts recovered as the 17-kDa species of IL-1β were multiplied by 2 to correct for loss due to proteolytic processing. Likewise, the quantity of radioactivity recovered intracellularly in association with the 31-kDa (31I) form and extracellularly in association with the 31-kDa (31e) and 17-kDa (17e) forms of IL-1α are indicated; counts recovered as the 17-kDa IL-1α species were multiplied by 1.5 to correct for loss due to proteolytic processing. Total radioactivity (sum of all individual species) also is indicated. Each value is the average of duplicate determinations.

Close Modal

or Create an Account

Close Modal
Close Modal