GLN is protective via PI3-K/Akt-Hsp70 signaling after HS. A) IEC-6 cells were treated with/without 1 h prior LY294002 (25 μM) treatment under non-stressed conditions (37°C). Cell viability was measured via MTS assay. Results are shown as mean ± SEM (n = 4). B) Cells were treated for 1 h with or without LY294002 (25 μM) before they were treated with 0, 2, or 10 mM GLN. Cell survival, following lethal HS (44°C), was measured via MTS assay. All groups were normalized to their non-HS controls to account for differences in cell growth. Assays were carried out in triplicate, experiments were performed 4 times and are shown as mean ± SEM. C) IEC-6 cells were treated with 0 mM or 10 mM GLN with or without LY294002 (25 μM) 1 h prior treatment under basal and stressed conditions (43°C). Hsp70 expression was determined by Western blot analysis (3 h recovery). In addition, ß-actin was measured to normalize total blotted protein. Data are shown as mean fold change relative to 0 mM GLN ± SEM (n = 4). D) Cells were treated as described in Figure 1C. Hsp70 expression is shown via ELISA in ng/ml ± SEM (n = 4).