Fig. 6. PTPL1-FERM interaction with PIPs. ³⁵S-labeled PTPL1-FERM domain (wt) and its KN1, KN2, KN1-2 mutants were produced by in vitro transcription-coupled translation system. (A) 4000 cpm of each labeled recombinant protein were analyzed on a 10% SDS/PAGE and submitted to autoradiography. (B) 40,000 cpm of each labeled protein were incubated with PIP-Strips (Echelon Biosciences) plotted with 100 pmol of PtdIns, PtdIns(3)P [PI(3)P], PtdIns(4)P [PI(4)P], PtdIns(5)P [PI(5)P], PtdIns(3,4)P₂ [PI(3,4)P2], PtdIns(3,5)P₂ [PI(3,5)P2], PtdIns(4,5)P₂ [PtdIns(4,5)P₂], PtdIns(3,4,5)P₃ [PI(3,4,5)P3], phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylserine (PS), lysophosphatidic acid (LPA), lysophosphacholine (LPC), sphingosine-1-phosphate (S1P) and phosphatic acid (PA). After three washes, PIP-strips were submitted to autoradiography.