Journal: Molecular and Cellular Biology

Article Title: Soluble Axl Is Generated by ADAM10-Dependent Cleavage and Associates with Gas6 in Mouse Serum

doi: 10.1128/mcb.25.21.9324-9339.2005

Figure Lengend Snippet: FIG. 2. Regulation of Axl shedding. L929 (A) and L929R (B) cells were treated with 200 ng/ml of PMA for different times, and the concentration of sAxl in cell-conditioned medium was evaluated by a specific ELISA. Untreated cells were used as controls. *, P 0.05 versus control samples. L929 (C) and L929R (D) cells were left untreated or treated with PMA for 2 h. Expression of membrane-bound Axl was evaluated by flow cytometry. (E and F) Cells were treated with PMA (200 ng/ml), Gas6 (50 ng/ml), IL-15 (50 ng/ml), TNF- (10 ng/ml), and LPS (10 ng/ml) for 2 h, and the concentration of sAxl in the culture medium was quantified by ELISA. Untreated (medium) cells were used as controls. **, P 0.01 versus control samples.

Article Snippet: Recombinant murine Gas6, rat antimouse Gas6, biotinylated goat anti-mouse Gas6, Axl, Mer, and Tyro3 (Dtk) Abs and Axl-Fc and IL-3R–Fc chimeras were purchased from R&D Systems.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Control, Expressing, Membrane, Cytometry

Journal: Molecular and Cellular Biology

Article Title: Soluble Axl Is Generated by ADAM10-Dependent Cleavage and Associates with Gas6 in Mouse Serum

doi: 10.1128/mcb.25.21.9324-9339.2005

Figure Lengend Snippet: FIG. 7. sAxl is present in mouse serum and associates with Gas6. (A) Sera from C57BL/6, CH3, BALB/c, Axl/, Axl/, and Axl/

Article Snippet: Recombinant murine Gas6, rat antimouse Gas6, biotinylated goat anti-mouse Gas6, Axl, Mer, and Tyro3 (Dtk) Abs and Axl-Fc and IL-3R–Fc chimeras were purchased from R&D Systems.

Techniques:

Journal: Molecular and Cellular Biology

Article Title: Soluble Axl Is Generated by ADAM10-Dependent Cleavage and Associates with Gas6 in Mouse Serum

doi: 10.1128/mcb.25.21.9324-9339.2005

Figure Lengend Snippet: FIG. 8. Immobilized Axl-Fc chimeric protein promotes cell migra- tion and induces the phosphorylation of Axl and PI3K. (A) L929R cells were plated onto Axl-Fc- or IL-3R–Fc (control)-coated six-well plates and allowed to grow in the presence of 10% FCS. After 18 h, a wound was created by scratching with a pipette tip (0 h). The cells were washed with PBS and incubated further to allow migration into the wounded area. Phase-contrast images of matched pairs of marked wound regions were taken 6 and 18 h later to assess cell migration. (B) L929R cells were serum starved for 4 h and incubated in Axl-Fc- coated wells for 15 min. Incubation in IL-3R–Fc (control)-coated wells or stimulation with 100 ng/ml of Gas6 was used as a negative or positive control, respectively. Cells were lysed, and Axl or PI3K was immunoprecipitated from the lysates with specific Abs. Precipitates were subjected to 10% SDS-PAGE and analyzed with anti-pTyr Abs. Detection of Axl or PI3K on the same blots was used as a loading control. The picture is representative of three independent experi- ments, all of which yielded highly comparable results. IP, immunopre- cipitation; WB, Western blotting.

Article Snippet: Recombinant murine Gas6, rat antimouse Gas6, biotinylated goat anti-mouse Gas6, Axl, Mer, and Tyro3 (Dtk) Abs and Axl-Fc and IL-3R–Fc chimeras were purchased from R&D Systems.

Techniques: Phospho-proteomics, Control, Transferring, Incubation, Migration, Positive Control, Immunoprecipitation, SDS Page, Western Blot

Journal: Molecular and Cellular Biology

Article Title: Soluble Axl Is Generated by ADAM10-Dependent Cleavage and Associates with Gas6 in Mouse Serum

doi: 10.1128/mcb.25.21.9324-9339.2005

Figure Lengend Snippet: FIG. 9. Immobilized Axl-Fc chimeric protein induces cell migration and phosphorylation of Axl and PI3K in WT but not Axl/ MEFs. (A) MEFs were plated onto Axl-Fc- or IL-3R–Fc (control)-coated six-well plates. A confluent cell monolayer was wounded by scratching with a pipette tip (0 h). The cells were washed with PBS and incubated further to allow migration into the wounded area. Phase-contrast images of matched pairs of marked wound regions were taken 6 and 18 h later to assess cell migration. (B) MEFs were serum starved for 4 h and incubated in Axl-Fc-coated wells for 15 min. Incubation in IL-3R–Fc-coated (control) wells or stimulation with 100 ng/ml of Gas6 was used as a negative or positive control, respectively. Cells were lysed, and Axl or PI3K was immunoprecipitated from the lysates with specific Abs. Precipitates were subjected to 10% SDS-PAGE and analyzed with anti-pTyr Abs. Detection of Axl or PI3K on the same blots was used as a loading control. IP, immunoprecipitation; WB, Western blotting.

Article Snippet: Recombinant murine Gas6, rat antimouse Gas6, biotinylated goat anti-mouse Gas6, Axl, Mer, and Tyro3 (Dtk) Abs and Axl-Fc and IL-3R–Fc chimeras were purchased from R&D Systems.

Techniques: Migration, Phospho-proteomics, Control, Transferring, Incubation, Positive Control, Immunoprecipitation, SDS Page, Western Blot