Fig. 4

PAF promotes cell migration via ADAM10 mediated activation of EGFR signaling pathways. a Fluorimetric ADAM10 activity assay performed on spreading SKCO15 model IECs treated with PAF (10 µM) or control at 30 min (*p < 0.5; n = 3; mean ± SEM). Immunoblotting was performed on lysates from scratch-wounded SKCO15 monolayers b and sparse primary IEC cultures c treated with PAF or control for 30 min. Levels of pEGFR, pSRC, and pFAK (Y397, Y861) were compared with total EGFR, SRC, FAK, and Calnexin to assess activation (representative blots from three experiments). c Confocal micrographs of the focal contacts in migrating IECs at the leading edge of the wound after 2-h treatment with control or PAF showing staining of pFAK (Y861), phalloidin (F-actin), and nuclei. (Scale bar 50 µm; original magnification ×40; representative images from 3 experiments). Statistical comparisons performed using unpaired two-tailed Student’s t test with Welch’s correction. NT vehicle control