Figure 8
Effect of acid and leptin on phosphorylation of FOXO1. Serum-starved OE33 cells were stimulated with either pH 4.0 acid or control bolus for 3 minutes followed by fresh serum-free medium with or without leptin (10 nM). Cells were formalin fixed 5 minutes after addition of leptin-containing medium for FOXO1 quantitation. Where indicated LY294004 10 μM was added 60 minutes before any stimulation. Phosphorylated and non-phosporylated total FOXO1 were quantified in formalin-fixed cells using ELISA assays. 8a: effect on total FOXO1.8b: effect on phsopho-FOXO1. Results expressed as mean ± SEM, N = 3. OD450 nm/OD 595 nm values represent specific reactivity of the peptide of interest in the ELISA corrected for the number of viable cells in the relevant well. * P < 0.05 vs untreated control, ** P < 0.05 vs acid or leptin alone, *** P < 0.05 vs stimulus in the absence of LY294004.