SPM Addition in cells

seed cells for 24h 0m 0s in a 10 cm dish at a density of 6 x 10⁶ cells per plate

treat cells for 24h 0m 0s with 10 µM freshly prepared SPM.

SPM (Sigma-Aldrich, Saint Louis, MO, USA) was prepared to a final stock concentration of 200 mM in 0.1 M 3-(N-morpholino)propane sulfonic acid (MOPS)-KOH (pH 7.0)

wash cells with 1X PBS and scrape cells for intact cell cross linking