Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Induction of potent TRAIL-mediated tumoricidal activity by hFLEX/Furin/TRAIL recombinant DNA construct.

doi: 10.1016/j.ymthe.2004.02.021

Figure Lengend Snippet: FIG. 2. Western blot analysis. (A) Expression of pFETZ-, pFFT-, and pFFZT-encoded proteins in COS7 cells following transfection. The presence of fusion proteins and cleaved protein products was detected using anti-TRAIL Ab. (B) Expression of pFFT, pFFZT, and pFETZ in vivo. pFFT-, pFFZT-, and pFETZ-encoded proteins could be detected in sera collected from mice injected with pFFT, pFFZT, or pFETZ with (i) anti-TRAIL Ab and (ii) anti-Flt3L Ab. (C) pFFT-, pFFZT-, or pFETZ- encoded proteins were consistently detected in sera collected from groups of mice injected with pFFT, pFFZT, or pFETZ, respectively. (D) Analysis of FFT, FFZT, and FETZ protein by 6% native PAGE. Cleaved form of TRAIL was assembled in stable trimers upon addition of isoleucine zipper sequence (arrow) compared to the monomeric cleaved TRAIL (dotted-line arrow).

Article Snippet: The membrane was then incubated with 5 Ag/ml rabbit anti-TRAIL polyclonal antibody or goat anti-Flt3L polyclonal antibody (Santa Cruz Biotechnology) for 1.5 h at room temperature.

Techniques: Western Blot, Expressing, Transfection, In Vivo, Injection, Clear Native PAGE, Sequencing

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Regression of human mammary adenocarcinoma by systemic administration of a recombinant gene encoding the hFlex-TRAIL fusion protein.

doi: 10.1006/mthe.2001.0280

Figure Lengend Snippet: FIG. 2. Western blot analysis of the fusion protein expressed in the BL6 cell line following transfection with the various recombinant DNA constructs. (A) Detection of hFlex, F81T, F81TZ, FET, and FETZ fusion proteins in the cell lysates by anti-Flt3L (left) and anti-TRAIL (right). (B) Presence of the FET and FETZ fusion proteins in cell culture medium. The F81T and F81TZ fusion proteins were not detectable in cell culture medium. (C) Analysis of FET and FETZ proteins by native 6% PAGE. Native FETZ was assembled in stable trimers compared to the monomeric FET.

Article Snippet: The membrane was then incubated with 5 mg/ml rabbit polyclonal anti-TRAIL antibody or goat polyclonal anti-Flt3L antibody (Santa Cruz Biotech, Santa Cruz, CA) for 1.5 h at room temperature.

Techniques: Western Blot, Transfection, Recombinant, Construct, Cell Culture

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Regression of human mammary adenocarcinoma by systemic administration of a recombinant gene encoding the hFlex-TRAIL fusion protein.

doi: 10.1006/mthe.2001.0280

Figure Lengend Snippet: FIG. 3. Determination of the concentration of the fusion proteins in Flt3L ELISA. (A) Cell medium was collected 24 h after transfection of BL6 cells with the recombinant genes and (B) serum was collected from mice 24 h after injection with 10 mg of recombinant genes. Each bar represents the mean and standard deviation of three independent samples.

Article Snippet: The membrane was then incubated with 5 mg/ml rabbit polyclonal anti-TRAIL antibody or goat polyclonal anti-Flt3L antibody (Santa Cruz Biotech, Santa Cruz, CA) for 1.5 h at room temperature.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Transfection, Recombinant, Injection, Standard Deviation

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Regression of human mammary adenocarcinoma by systemic administration of a recombinant gene encoding the hFlex-TRAIL fusion protein.

doi: 10.1006/mthe.2001.0280

Figure Lengend Snippet: FIG. 4. Time course of expression of the fusion protein following a single injection of different amounts of pFETZ plasmid DNA in SCID mice. The level of FETZ in the serum was determined using Flt3L ELISA. Each time point represents the mean and standard deviation of three mice.

Article Snippet: The membrane was then incubated with 5 mg/ml rabbit polyclonal anti-TRAIL antibody or goat polyclonal anti-Flt3L antibody (Santa Cruz Biotech, Santa Cruz, CA) for 1.5 h at room temperature.

Techniques: Expressing, Injection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Regression of human mammary adenocarcinoma by systemic administration of a recombinant gene encoding the hFlex-TRAIL fusion protein.

doi: 10.1006/mthe.2001.0280

Figure Lengend Snippet: FIG. 5. Ability of FETZ to induce cell killing of human mammary adenocar- cinoma MDA-231 cells. Cells were incubated with serum, containing fusion proteins (1 mg/ml), obtained from mice 12 h following injection with the various recombinant TRAIL DNA constructs. The concentration of the fusion protein was determined by Flt3L ELISA. Commercial human recombinant TRAIL (hTrail) protein was diluted in normal mouse serum.

Article Snippet: The membrane was then incubated with 5 mg/ml rabbit polyclonal anti-TRAIL antibody or goat polyclonal anti-Flt3L antibody (Santa Cruz Biotech, Santa Cruz, CA) for 1.5 h at room temperature.

Techniques: Incubation, Injection, Recombinant, Construct, Concentration Assay, Enzyme-linked Immunosorbent Assay