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| | Diabetes: Glycogen Synthase Sensitivity to Insulin and Gl... @ HighBeam Research |
 | | We conclude that Ser-7 and Ser-640 are both involved in mediating the response of glycogen synthase to insulin and activation by glucose 6-phosphate. |
 | | Stable clones of Rat-1 fibroblasts were cultured to confluent monolayers in 60-mm dishes in DMEM with 25 mmol/l glucose supplemented with 10% FBS, 100 U/ml penicillin, 100 [micro]g/ml streptomycin, and 0.4 mg/ml G418, followed by 48-h incubation in the same media with 5 mmol/l glucose. |
 | | After incubation for 1 h, cells were rinsed with 6 ml of ice-cold homogenization buffer (HB) (50 mmol/l Tris-HCl, pH 7.8, 10 mmol/l EDTA, 2 mmol/l EGTA, 100 mmol/lNaF, 0.1 mmol/l [N.sup.[Alpha]]-p-tosyl-L-lysine chloromethyl ketone, 1 mmol/l phenylmethylsulfonyl fluoride, and 1 mmol/l dithiothreitol) and were frozen in liquid nitrogen in 0.5 ml of HB buffer. |
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