Fig. 7.

Detection of NHE1, Hsp70, and GSH after immunoprecipitation with anti-NHE1. Monocytes were incubated with the inhibitors for 15 min/37 °C followed by incubation with leptin (16 ng/ml) for 30 min/37 °C and then lysed. Cell extracts were immunoprecipitated with anti-NHE1 and then analyzed through Western blot. Anti-Hsp70 and anti-GSH blots were observed at the same molecular weight marker (approximately 70 kDa) indicating that Hsp70 was glutathionylated. Samples: 1 control, 2 leptin 16 ng/ml, 3 leptin 16 ng/ml + cariporide 20 nM, 4 leptin 16 ng/ml + wortmannin 50 nM, 5 leptin 16 ng/ml + GF109203X 10 μΜ