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S1P antibodies decrease S1P- and 4-Deoxypyridoxine-induced HPAEC migration.

By Evgeny V. Berdyshev (307465), Irina Gorshkova (356721), Peter Usatyuk (356723), Satish Kalari (307462), Yutong Zhao (356724), Nigel J. Pyne (356726), Susan Pyne (356727), Roger A. Sabbadini (356730), Joe G. N. Garcia (118950) and Viswanathan Natarajan (307479)


<p>HPAECs (∼90% confluence) grown on gold electrodes or coverslips were starved for 3 h in 0.1% FBS in EBM-2 without growth factors. <b><i>A</i></b> -Cells were treated with 4-DP (1 mM for 30 min) then wounded on the gold electrodes as described under “Experimental Procedures” prior to the addition of isotype-matched control IgGk1 or anti-S1P antibody (150 µg/ml) and S1P (1 µM) challenge. Transendothelial electrical resistance was recorded for 16 h. Values are the mean ± S.E.M. for three independent experiments each performed in triplicate. <b><i>B,C</i></b> - Cells on cover slips were treated with 4-DP (1 mM) and anti-S1P antibody for 6 h, then stimulated with 0.1% BSA-complexed S1P (1.0 µM for 5 min), washed, fixed, permeabilized, probed with anti-Rac1 antibody (<b><i>B</i></b>) or anti-IQGAP1 antibody (<b><i>C</i></b>), and examined by immunofluorescence microscopy using a ×60 oil objective. Shown is a representative micrograph from several independent experiments.</p

Topics: Biochemistry, antibodies, s1p-, 4-deoxypyridoxine-induced, hpaec
Year: 2013
DOI identifier: 10.1371/journal.pone.0016571.g013
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Provided by: FigShare
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