Silica gel 60a thin layer chromatography plates

The HEK293 (immortalized human embryonic kidney cells), HN33 (immortalized rat hippocampal neuronal cells), and U87MG (immortalized human glioblastoma cells) cell lines were cultured at 37°C in Dulbecco's modified Eagle’s medium (DMEM, Gibco-BRL, CA, USA) containing 10% fetal bovine serum (FBS) and 1% antibiotic-antimycotic solution. The U87MG cells stably overexpressing the WT or the catalytically inactive mutant (K758R) of PLD2 (PLD2-KRm) were used as reported previously (26 (link)). PLD2-PH-expressing U87MG cells were esta-blished by transfection using Lipofectamine and selected using G418 (700 μg/ml). Protein A-Sepharose and Glutathione Sepharose 4B were obtained from Amersham Biosciences (UK). The phosphatidylbutanol (PtdBut) standard was acquired from Avanti Polar Lipids, Inc. (AL, USA). [9,10-³H]myristic acid was purchased from Perkin-Elmer Life Sciences (MA, USA). The silica gel 60A thin-layer chromatography plates were obtained from Whatman (UK). The ECL Western Blotting Detection Kit was procured from Amersham Biosciences (UK).