Journal: World Journal of Gastroenterology

Article Title: Fas counterattack in cholangiocarcinoma: A mechanism for immune evasion in human hilar cholangiocarcinomas

doi: 10.3748/wjg.v7.i6.860

Figure Lengend Snippet: FasL positive in human hilar cholangiocarcinom as (brown). SABC × 200

Article Snippet: Immunohistochemistry for FasL and CD45 Detection of FasL expression and CD45 positive cells was performed using a rabbit polyclonal anti-human FasL specific IgG and a mouse anti-human monoclonal antibody on paraffin sections of human cholangiocarcinoma respectively (Boster Biological Technology Company, Wuhan, China).

Techniques:

Journal: World Journal of Gastroenterology

Article Title: Fas counterattack in cholangiocarcinoma: A mechanism for immune evasion in human hilar cholangiocarcinomas

doi: 10.3748/wjg.v7.i6.860

Figure Lengend Snippet: Expression of FasL in cholangiocarcinoma cell line. QBC939 × 200

Article Snippet: Immunohistochemistry for FasL and CD45 Detection of FasL expression and CD45 positive cells was performed using a rabbit polyclonal anti-human FasL specific IgG and a mouse anti-human monoclonal antibody on paraffin sections of human cholangiocarcinoma respectively (Boster Biological Technology Company, Wuhan, China).

Techniques: Expressing

Journal: World Journal of Gastroenterology

Article Title: Fas counterattack in cholangiocarcinoma: A mechanism for immune evasion in human hilar cholangiocarcinomas

doi: 10.3748/wjg.v7.i6.860

Figure Lengend Snippet: Expression of FasL mRNA in human cholangiocarcinoma cells QBC939. M: DL 2000 Marker; 1: FasL; 2: FasL+β-actin

Article Snippet: Immunohistochemistry for FasL and CD45 Detection of FasL expression and CD45 positive cells was performed using a rabbit polyclonal anti-human FasL specific IgG and a mouse anti-human monoclonal antibody on paraffin sections of human cholangiocarcinoma respectively (Boster Biological Technology Company, Wuhan, China).

Techniques: Expressing, Marker

Journal: World Journal of Gastroenterology

Article Title: Fas counterattack in cholangiocarcinoma: A mechanism for immune evasion in human hilar cholangiocarcinomas

doi: 10.3748/wjg.v7.i6.860

Figure Lengend Snippet: Western blotting of FasL protein with mAb from QBC939 cell cultures clone 33 from QBC939 cell cultures

Article Snippet: Immunohistochemistry for FasL and CD45 Detection of FasL expression and CD45 positive cells was performed using a rabbit polyclonal anti-human FasL specific IgG and a mouse anti-human monoclonal antibody on paraffin sections of human cholangiocarcinoma respectively (Boster Biological Technology Company, Wuhan, China).

Techniques: Western Blot

Journal: Membranes

Article Title: Breakdown of Phospholipid Asymmetry Triggers ADAM17-Mediated Rescue Events in Cells Undergoing Apoptosis

doi: 10.3390/membranes13080720

Figure Lengend Snippet: Caspase-3 activation induces increased shedding of ADAM17 substrates in Xkr8 overexpressing cells. ( A ) Caspase-3 is similarily activated in HEK293T cells transfected with Xkr8 or mock vector upon stimulation with kTRAIL (50 ng/mL) and CHX (5 µg/mL) and reduced in the presence of pan-caspase inhibitor ZVAD (50 µM). Caspase-3 activity of one representative experiment over time. ( B ) Quantification of caspase-3 activity experiments ( n = 4; */# p < 0.05; ±SEM). ( C , D ) HEK293T cells were co-transfected with Xkr8 or mock vector and AP-tagged ADAM17 substrates TGF-alpha ( C ) or epiregulin ( D ), respectively. Cells were stimulated CHX (5 µg/mL) alone (mock) or with kTRAIL (50 ng/mL) and CHX (5 µg/mL) overnight in the presence of absence of caspase inhibitor ZVAD (50 µM). Constitutive and apoptosis-induced shedding was significantly increased upon overexpression of Xkr8. * Significant increase compared with indicated control, # significant decrease compared with respective kTRAIL-treated cells. ( C , D ) n = 3; */# p < 0.05; ±SEM).

Article Snippet: Anti-ADAM17 antibodies used for Western blotting were from Merck Millipore (Darmstadt, Germany; AB19027) and from Cell Signaling Technology (3976S).

Techniques: Activation Assay, Transfection, Plasmid Preparation, Activity Assay, Over Expression

Journal: Membranes

Article Title: Breakdown of Phospholipid Asymmetry Triggers ADAM17-Mediated Rescue Events in Cells Undergoing Apoptosis

doi: 10.3390/membranes13080720

Figure Lengend Snippet: PMA-stimulation leads to increased shedding of TGF-alpha and EREG in Xkr8 overexpressing cells. HEK293T cells were co-transfected with mock vector or Xkr8 and ( A ) AP-tagged TGF-alpha or ( B ) EREG, respectively. Cells were stimulated with PMA (100 ng/mL) for 30 min and analyzed for substrate shedding. ( C , D ) HEK293T cells were analyzed for TGF-alpha ( C ) or EREG ( D ) shedding after 30 min in the presence or absence of broad-spectrum metalloprotease inhibitor marimastat (MM, 10 µM) or OPS. ( E , F ) Cells were stimulated with PMA (100 ng/mL) for 30 min in the presence or absence of marimastat (MM, 10 µM), ADAM10 inhibitor GI (3 µM), or the ADAM17/ADAM10 inhibitor GW (3 µM) and analyzed for substrate shedding. ( G , H ) Cells were stimulated with PMA (100 ng/mL) for 30 min in the presence or absence of marimastat (MM, 10 µM)) or the ADAM17 inhibitory antibody D1 (200 nM) and analyzed for substrate shedding. * significant increase compared with indicated control, # significant decrease compared with corresponding stimulated cells. ( A – D , E , G ): n ≥ 4; ±SEM. ( F , H ): n = 3 ± SEM; */# p < 0.05).

Article Snippet: Anti-ADAM17 antibodies used for Western blotting were from Merck Millipore (Darmstadt, Germany; AB19027) and from Cell Signaling Technology (3976S).

Techniques: Transfection, Plasmid Preparation

Journal: Membranes

Article Title: Breakdown of Phospholipid Asymmetry Triggers ADAM17-Mediated Rescue Events in Cells Undergoing Apoptosis

doi: 10.3390/membranes13080720

Figure Lengend Snippet: Decreased expression of Xkr8 leads to decreased release of soluble EREG and decreased cellular ATP content. ( A ) Caco-2 cells were either transfected with control siRNA (siCtr) or with Xkr8 siRNA (siXkr8). After 48 h, cells were stimulated with CHX (5 µg/mL, mock) or with kTRAIL (100 ng/mL) and CHX in the absence or presence of ZVAD (50 µM) and analyzed for release of soluble EREG via ELISA. ( B ) Caco-2 cells were either transfected with control siRNA or with Xkr8 siRNA and either mock stimulated or stimulated with kTRAIL/CHX. Mock-transfected cells were additionally incubated in the presence of ADAM10/ADAM17 inhibitor GW (3 µM). After overnight incubation, ATP was measured using a luminometric ATP cell viability assay. The kTRAIL-treated cells are compared with the respective non-treated controls, which were set to 100 percent ( n = 3; * p < 0.05; ±SEM). * significant increase compared with respective control, # significant decrease compared with corresponding cells. ( C ) Proposed model of Xkr8-dependent ADAM activation. Caspase activation induces phospholipid scrambling via Xkr8 or related scramblases. PS exposure activates ADAM17 sheddase function and release of growth factors that might counteract the death cascade.

Article Snippet: Anti-ADAM17 antibodies used for Western blotting were from Merck Millipore (Darmstadt, Germany; AB19027) and from Cell Signaling Technology (3976S).

Techniques: Expressing, Transfection, Enzyme-linked Immunosorbent Assay, Incubation, Viability Assay, Activation Assay