Figure 10. GOP-1 promotes loading of GTP... | Rockefeller University Press

Figure 10.

$Figure 10. GOP-1 promotes loading of GTP onto and membrane recruitment of UNC-108. GOP-1 promotes association of UNC-108 with liposomes. Binding of UNC-108–GDI-1 to PC + PtdIns3P (8%) liposomes was detected by Western blot. UNC-108 but not GDI-1 was recovered in the top fraction with GOP-1 in a liposome flotation assay in the absence or presence of GTPγS. Quantification is shown at right. At least three independent experiments were performed. Data are shown as mean ± SD. Unpaired t test was performed to compare datasets linked by lines. *, P < 0.05. (B) GOP-1 promotes loading of GTP onto UNC-108. Schematic illustration of the GTP incorporation assay is shown at left. At least three independent experiments were performed, and a representative result is shown. (C–H) Fluorescent images of the gonads (C and D) or coelomocytes (F and G) in gop-1(tm5384) expressing GFP::UNC-108 (C and F) or GFP::UNC-108(K120E) (D and G). K120E (white arrowheads) but not wild-type UNC-108 (yellow arrowheads) was seen on phagosomes (C and D) and endosomes (F and G). Quantification is shown in E and H. In E, at least 15 animals were scored, and data are shown as mean ± SD. Unpaired t test was performed to compare data derived from wild type and gop-1(tm5384). **, P < 0.0001; N.S., no significant difference. In H, the percentage of animals containing GFP-positive vacuoles in coelomocytes was quantified. n, number of animals followed and quantified. Bars, 5 µm.$

GOP-1 promotes loading of GTP onto and membrane recruitment of UNC-108. GOP-1 promotes association of UNC-108 with liposomes. Binding of UNC-108–GDI-1 to PC + PtdIns3P (8%) liposomes was detected by Western blot. UNC-108 but not GDI-1 was recovered in the top fraction with GOP-1 in a liposome flotation assay in the absence or presence of GTPγS. Quantification is shown at right. At least three independent experiments were performed. Data are shown as mean ± SD. Unpaired t test was performed to compare datasets linked by lines. *, P < 0.05. (B) GOP-1 promotes loading of GTP onto UNC-108. Schematic illustration of the GTP incorporation assay is shown at left. At least three independent experiments were performed, and a representative result is shown. (C–H) Fluorescent images of the gonads (C and D) or coelomocytes (F and G) in gop-1(tm5384) expressing GFP::UNC-108 (C and F) or GFP::UNC-108(K120E) (D and G). K120E (white arrowheads) but not wild-type UNC-108 (yellow arrowheads) was seen on phagosomes (C and D) and endosomes (F and G). Quantification is shown in E and H. In E, at least 15 animals were scored, and data are shown as mean ± SD. Unpaired t test was performed to compare data derived from wild type and gop-1(tm5384). **, P < 0.0001; N.S., no significant difference. In H, the percentage of animals containing GFP-positive vacuoles in coelomocytes was quantified. n, number of animals followed and quantified. Bars, 5 µm.