Journal: Cell Death & Disease

Article Title: Biliary tract instillation of a SMAC mimetic induces TRAIL-dependent acute sclerosing cholangitis-like injury in mice

doi: 10.1038/cddis.2016.459

Figure Lengend Snippet: cIAP-1 and cIAP-2 are downregulated in cholangiocytes of PSC patients. ( a ) Representative images of liver sections stained for cIAP-1 (left panel) and cIAP-2 (right panel) in patients with normal, NASH and PSC (stage IV) liver histology. Photomicrographs of small bile ducts (SBD) and large bile ducts (LBD) taken at × 20 magnification. The arrows point to the bile ducts. ( b ) Histological scoring for cIAP-1 in normal (5, 31), NASH (12, 120) and PSC (16, 131) patients and for cIAP-2 in normal (5, 31), NASH (12, 118) and PSC (16, 123) patients. Numbers in parentheses indicate total number of patients and total number of small and large bile ducts evaluated, respectively. Grade 0=no protein expression; grade 3=high protein expression. ** P <0.01, *** P <0.001. ( c ) Representative images of liver sections stained for TWEAK (left panel) and Fn14 (right panel) from patients with normal or PSC (stage IV) liver histology. Photomicrographs of small bile ducts and large bile ducts taken at × 40 and × 20 magnification, respectively. The arrows point to the bile ducts. ( d ) Immunoblot analysis showing expression of cIAP-1, cIAP-2 and actin (loading control) in H69 cells treated with human recombinant TWEAK (100 ng/ml) for the indicated times

Article Snippet: Human recombinant TWEAK was from R&D Systems.

Techniques: Staining, Expressing, Western Blot, Recombinant

Journal: PLoS ONE

Article Title: Fn14•Trail Effectively Inhibits Hepatocellular Carcinoma Growth

doi: 10.1371/journal.pone.0077050

Figure Lengend Snippet: (A) The amino-acid sequence of the Fn14-TRAIL protein. The amino-acid sequence of the extra-cellular domain of human Fn14 (amino-acids 1-52 of the mature protein, marked in bold letters) are directly linked to the extra-cellular domain of human TRAIL (amino-acids 53-217 of the mature protein, non-bold letters). The underlined sequence represents the signal-peptide of the human Urokinase protein, utilized to secrete Fn14-TRAIL out of the cell and removed from the mature protein. (B) Fn14-TRAIL separated at denaturizing conditions on SDS-PAGE, Coomassie gel staining. (C) Western blot analysis with anti-TRAIL and anti-Fn14 primary antibodies.

Article Snippet: Soluble Fn14 (recombinant human Tweak receptor, PeproTech) was used instead of Fn14-Fc in some experiments.

Techniques: Sequencing, SDS Page, Staining, Western Blot

Journal: PLoS ONE

Article Title: Fn14•Trail Effectively Inhibits Hepatocellular Carcinoma Growth

doi: 10.1371/journal.pone.0077050

Figure Lengend Snippet: ( A ) The mRNA expression level of TRAIL, TRAIL receptors (DR4, DR5, DCR-1, DCR-2, OPG), Fn14 and TWEAK was determined by quantitive real-time PCR analysis. A representative experiment of three independent experiments is shown. Data are shown as average of triplicates (SD < 0.3), normalized against two endogenous control human genes, TBP and Actin-B, as calculated by Dataassist v2.0 software. ( B , C ) Protein expression of TRAIL, TRAIL receptors (DR4, DR5, DcR1, DcR2), Fn14 and TWEAK was determined by flow cytometeric analysis. ( D ) Fn14•TRAIL binds to HCC cells – HepG2 cells were incubated with Fn14•TRAIL, soluble TRAIL, Fn14 or the combination of the latter for 30 min at 4°c, immune-stained with PE-conjugated anti-Fn14, and analyzed by flow-cytometry. The results represent the mean +/- SD of triplicates (* p ≤ 0.05).

Article Snippet: Soluble Fn14 (recombinant human Tweak receptor, PeproTech) was used instead of Fn14-Fc in some experiments.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Software, Incubation, Staining, Flow Cytometry

Journal: PLoS ONE

Article Title: Fn14•Trail Effectively Inhibits Hepatocellular Carcinoma Growth

doi: 10.1371/journal.pone.0077050

Figure Lengend Snippet: ( A ) SK-HEP-1 [A, left panel], HepG2 [A, middle panel] and Huh7 [A, right panel] HCC cell lines, as well as NKNT3 [B, left panel] and FHB [B, right panel] hepatocyte cell lines, were incubated with 0, 3, 30 or 300 ng/ml of Fn14•TRAIL, TRAIL, Fn14-Fc or combination of the later two for 48 hours. Viable cells were stained with trypan blue and counted. The results represent the mean +/- SD of three independent experiments (* p ≤ 0.05). ( B ) HepG2 HCC cells were incubated with 30ng/ml Fn14•TRAIL for 24 hours, in the presence or absence of anti Fn14 or anti TRAIL blocking antibodies. Treated cells were stained by Annexin V-FITC and Propidium Iodide, and counted by flow cytometer (2x10 4 cells per sample). The results represent the mean +/- SD of two independent experiments (* p ≤ 0.05).

Article Snippet: Soluble Fn14 (recombinant human Tweak receptor, PeproTech) was used instead of Fn14-Fc in some experiments.

Techniques: Incubation, Staining, Blocking Assay, Flow Cytometry