FIG.4. Requirement of the band 4.1/FERM domain for the subcellular localization of PTP-FERM. (A) Schematic drawings of PTP-FERM mutants used in the study. The solid lines represent the portions contained in the mutants. Amino acid numbers are shown below the lines. (B)Subcellular localization of PTP-FERM and mutants in the worm. DNA constructs containing ptp-ferm::ptp-ferm-gfp and its mutants were injected into the gonad of worm and lines that had these DNA constructs as extrachromosomal arrays were established. a to c, PTP-FERM-GFP; d to f,
del4.1; g to i,
del4.1delPDZ; j to l, delPDZ; m to o, delPTP; p to r, 4.1. Images of the neurons in the ventral nerve cord (a, c, d, f, g, i, j, l, m, o, p, and r) or on the lateral side of the worm (b, e, h, k, n, and q) are shown. The magnificationof c, f, i, l, o, and r is twofold that of others. Large and small arrows indicate nerve processes and cell bodies, respectively. Scale bars, 5 um. (C) Confocal microscopy analysis of PTP-FERM in mammalian 293T cells. 293Tcells were transiently transfected with control vector (a) or a plasmid coding for PTP-FERM-Flag (b) or Flag-PTP-FERM (c) for two days. After fixation and permeabilization, PTP-FERM was detected using anti-Flag antibody and FITC-conjugated goat anti-mouse IgG. Cells were visualized by confocal microscopy. Scale bar, 5 um. (D) The necessity of the FERM domain for the membrane localization of PTP-FERM in mammalian 293Tcells. PTP-FERM-GFP and its deletion mutants were transiently expressed in 293T cells. a, PTP-FERM-GFP; b,
del4.1; c,
del4.1delPDZ; d, delPDZ;e, delPTP; f, 4.1; g, 4.1del1; h, 4.1del2. The pattern of GFP alone is indistinguishable from that of 4.1del1 and 4.1del2 (data not shown). Scale bar, 5 um.