“…The cells were washed with PBS and lysed with 1 ml of 10% trichloracetic acid (TCA). The lysates were centrifuged and the supernatants were extracted five times with two volumes of water-saturated diethyl ether and neutralized by the addition of SO p1 0.3 M EDTA and 9.6 p1 0.6 M NaHCO, (Munoz-Acedo and Izquierdo-Claros, 1995). The samples were then applied to columns containing 1 ml of Bio-Rad AG 1-X8 anion exchange resin (formatc form, 200-400 mesh), and inositol phosphates were separated by stepwise elution, as described previously (Joseph et al, 1989).…”