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Fig. 4. The localization of the PTPL1-FERM domain is altered by mutation of the two potential PtdIns(4,5)P2-binding sites in COS1 cells. COS1 cells were transfected with (A) HA-epitope tagged wild-type FERM domain, (B) KN1-FERM mutant (K645, 646, 647N), (C) KN2-FERM mutant (K824, 825, 830, 831, 832N) and (D) KN1-2-FERM mutant (combination of KN1 and KN2 mutations) and treated for immunolocalization of HA-epitope tagged constructs as described in Fig. 2. (A-C) Transfected wild-type FERM domain, KN1 and KN2 mutants were associated with the plasma membrane, dorsal microvilli and induced membrane extensions (arrowheads). (D) KN1-2 mutant of FERM domain was mainly localized in the cytoplasm (arrowhead). Bars, 10 µm.