GRGDSP’s effect on cell area size, F-actin morphology and intracellular GLN concentration.

<p>A) Cell area size is shown by fluorescence microscopy in IEC-6 cells under 0 mM GLN conditions and 10 mM GLN treatment without inhibitor G<u>RGD</u>SP (Images A, C, E, G) or after 1 h prior treatment with G<u>RGD</u>SP (Images C, D, F, H) in basal and stressed conditions (43°C). F-actin is shown in green and nuclei in blue. All images taken 15 min following exposure of cells to GLN in non-HS cells and 15 min post-HS in stressed cells. All images were aquired using the same exposure times, and were renormalized the same for optical comparison (n = 4). Scalebars = 33 µm. B) Cell area size data are shown in fold increase under non-HS (37°C) and HS (43°C) conditions (n = 3–4) C) IEC-6 cells were treated as descibed in A). GLN levels in cell extracts were quantified using LC-MS/MS and are shown as fold change± SEM ratioed to 0 mM GLN group (n = 6–7). D) Fluorescence microscopy was used for the morphological analysis of the distribution of F-actin. Microfilaments were visualized with Alexa Flour 488 phalloidin antibody. Representative results are shown from 4 independent fluorescence microscopy experiments for each condition. Scalebars: 45 µm.