TABLE 3 

Composition of the buffers and cell culture media used in this study

Buffer or mediumComposition
Kinase IP20 mM Tris (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% Triton X-100, 2.5 mM sodium pyrophosphate, 1 mM β-glycerophosphate, 1 mM Na3VO4, 1 mM dithiothreitol (DTT), 0.5% NP-40, 2 cOmplete mini EDTA-free protease inhibitor tablets (Roche), 5 μl of Benzonase nuclease/10 ml
MBP lysis/gel filtration500 mM NaCl, 50 mM CAPS,a 5 mM DTT, 10% glycerol (pH 10)
GST lysis/gel filtration300 mM NaCl, 50 mM Na2HPO4, 5 mM DTT, 5% glycerol (pH 7.4)
His lysis/gel filtration50 mM Tris (pH 8), 150 mM NaCl, 10% glycerol, 2 mM DTT
ADP-ribosylation50 mM Tris (pH 7.4), 1 mM DTT, 60 μM ATP, 5 mM MgCl2
1× Kinase buffer100 mM KCl, 100 mM MOPS,b 10 mM DTT, 10 mM MgCl2, 100 µM ATP
Enterocyte dissociation buffer1× Hanks’ balanced salt solution without Mg and Ca plus 10 mM HEPES, 1 mM EDTA, and 5 µl/ml 2-β-mercaptoethanol
HeLa, A549, or Caco2 cell maintenanceDulbecco’s modified Eagle’s medium (Sigma-Aldrich) containing 1,000, 1,000, or 4,500 mg/liter glucose, 1% (vol/vol) GlutaMAX (Life Technologies, Inc.), and 10% (HeLa, A549) or 15% heat-inactivated FCS
Thp1 cell maintenanceRPMI supplemented with 10% FCS
  • a CAPS, N-cyclohexyl-3-aminopropanesulfonic acid.

  • b MOPS, morpholinepropanesulfonic acid.