Brain l α phosphatidylinositol 4 5 bisphosphate

Natural and synthetic phospholipids, including POPS, POPC, POPE_, brain total lipid extract, brain L-α-phosphatidylinositol-4,5-bisphosphate, and TopFluor-TMR-PI(4,5)P₂ are from Avanti Polar Lipids. OG-DHPE and β-casein from bovine milk (>98% pure) were from Sigma-Aldrich. Alexa Fluor 647 and 488 Maleimide labeling kits are from Invitrogen. Culture-Inserts 2 Well for self-insertion were purchased from ibidi.
The following antibodies were used in this study: monoclonal mouse antibody to BIN1 (clone C99D against exon 17) from Millipore, polyclonal rabbit antibody to IRSp53 (BAIAP2) from Atlas Antibodies (HPA023310), HRP-conjugated beta-actin monoclonal antibody from Proteintech (HRP-60008), ezrin antibody from M. Arpin laboratory^{75 (link)}, monoclonal rabbit antibody to phospho-Ezrin (Thr567)/Radixin (Thr564)/Moesin (Thr558) Cell Signaling (Cat. 3726). Atto390 phalloidin was from Sigma.

1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] (DGS-NTA), 1,2-dioleoyl-sn-glycero-3-phosphocholine (PC), 1,2-dioleoyl-sn-glycero-3-phospho-l-serine (PS), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (PE), brain L-α-phosphatidylinositol-4,5-bisphosphate (PI), and E.coli cardiolipin (CL) were obtained from Avanti Polar Lipids. Nitrilotriacetic acid (Ni²⁺-NTA)-chelating Sepharose and Superdex 200 increase 10/300 GL were purchased from GE Healthcare. All other chemicals were acquired from Sigma.