Journal: British Journal of Cancer

Article Title: Antiapoptotic effect of haem oxygenase-1 induced by nitric oxide in experimental solid tumour

doi: 10.1038/sj.bjc.6600830

Figure Lengend Snippet: Apoptosis induction and change in HO activity after treatment with ZnPP IX of AH136B solid. Apoptosis ( A, B ) and HO activity ( C ) were assessed on day 14 after tumour implantation. Each specimen was analysed via TUNEL staining 24 h after treatment with vehicle (the control) ( A, a ), with ZnPP IX (500 μ g kg −1 i.a.) ( A, b ), or with CuPP IX (500 μ g kg −1 i.a.) ( A, c ). Quantitative analysis of TUNEL-positive cells in each specimen is shown ( B ). TUNEL-positive cells were counted in four different fields of magnification at × 100 per sample, and then the number of positive cells per mm 2 was calculated. Haem oxygenase activity of the solid tumour was measured after treatment in the same manner as in the TUNEL analysis ( C ). ** P <0.05, ** P <0.01 vs control ( n =3 for each group). Data are means±s.e. See text for details.

Article Snippet: AH136B tumour (rat hepatoma) cells were implanted subcutaneously (s.c.) in a dorsal site on the foot of male Donryu rats weighing 160–180 g (SLC, Inc., Shizuoka, Japan) with an inoculum size of 1 × 10 7 cells per injection site as described previously ( Doi et al , 1996 ).

Techniques: Activity Assay, TUNEL Assay, Staining, Control

Journal: British Journal of Cancer

Article Title: Antiapoptotic effect of haem oxygenase-1 induced by nitric oxide in experimental solid tumour

doi: 10.1038/sj.bjc.6600830

Figure Lengend Snippet: Zinc protoporphyrin IX-induced apoptosis of AH136B cells in vitro . AH136B cells were incubated for 24 h with indicated concentrations of ZnPP IX, with or without caspase-3 inhibitor ( A ) or SnPP IX ( B ), or with 100 μ M CuPP IX. ( A, B ) TUNEL-positive cells were counted in four different fields of magnification at × 100 per sample, and then the percentage of positive cells was calculated. ** P <0.05 vs control ( n =3 for each group). Cell viability after the same treatment is shown in the inset. ** P < 0.05, ** P <0.01 vs control ( n =4 for each group). Data are means±s.e. ( C ) Representative TUNEL staining of control and ZnPP IX-treated tumour cells. See text for details.

Article Snippet: AH136B tumour (rat hepatoma) cells were implanted subcutaneously (s.c.) in a dorsal site on the foot of male Donryu rats weighing 160–180 g (SLC, Inc., Shizuoka, Japan) with an inoculum size of 1 × 10 7 cells per injection site as described previously ( Doi et al , 1996 ).

Techniques: In Vitro, Incubation, TUNEL Assay, Control, Staining

Journal: British Journal of Cancer

Article Title: Antiapoptotic effect of haem oxygenase-1 induced by nitric oxide in experimental solid tumour

doi: 10.1038/sj.bjc.6600830

Figure Lengend Snippet: Effect of ZnPP IX on HO activity and caspase-3 activity of AH136B cells cultured in vitro . Haem oxygenase activity was measured 24 h after incubation with vehicle (control), ZnPP IX, or CuPP IX ( A ). Similarly, cells were treated with 100 μ M ZnPP with or without caspase-3 inhibitor (1 or 10 μ M ) for 24 h ( B ). Caspase-3 activity was measured fluorome-trically in cell extracts by using a fluorescent substrate. ** P <0.05, ** P <0.01 vs control ( n =3 for each group). Data are means±s.e. See text for details.

Article Snippet: AH136B tumour (rat hepatoma) cells were implanted subcutaneously (s.c.) in a dorsal site on the foot of male Donryu rats weighing 160–180 g (SLC, Inc., Shizuoka, Japan) with an inoculum size of 1 × 10 7 cells per injection site as described previously ( Doi et al , 1996 ).

Techniques: Activity Assay, Cell Culture, In Vitro, Incubation, Control

Journal: British Journal of Cancer

Article Title: Antiapoptotic effect of haem oxygenase-1 induced by nitric oxide in experimental solid tumour

doi: 10.1038/sj.bjc.6600830

Figure Lengend Snippet: Protective effect of bilirubin against ZnPP IX-induced apoptosis of AH136B cells. AH136B cells were incubated for 24 h with 100 μ M ZnPP IX in the presence or absence of indicated concentrations of bilirubin. TUNEL-positive cells were counted in four different fields of magnification at × 100 per sample, and then the percentage of positive cells was calculated. ** P <0.05 vs ZnPP IX alone ( n =3 for each group). Data are means±s.e. See text for details.

Article Snippet: AH136B tumour (rat hepatoma) cells were implanted subcutaneously (s.c.) in a dorsal site on the foot of male Donryu rats weighing 160–180 g (SLC, Inc., Shizuoka, Japan) with an inoculum size of 1 × 10 7 cells per injection site as described previously ( Doi et al , 1996 ).

Techniques: Incubation, TUNEL Assay

Journal: British Journal of Cancer

Article Title: Antiapoptotic effect of haem oxygenase-1 induced by nitric oxide in experimental solid tumour

doi: 10.1038/sj.bjc.6600830

Figure Lengend Snippet: Tumour blood flow after ZnPP IX or L -NAME treatment. Tumour blood flow was measured in AH136B solid tumour on day 14 after tumour implantation. At the indicated times after injection of 500 μ g kg −1 ZnPP IX (i.a.) or 6 mg kg −1 L -NAME (i.p.), a laser Doppler flowmeter and a probe needle were used to measure blood flow. Relative changes in tumour blood flow after injection of ZnPP IX or L -NAME are shown. Inset, change in tumour blood flow during early period after treatment (within 25 min). ** P <0.01 by ANOVA between the groups ( n =3 for each time point). Data are means±s.e. See text for details.

Article Snippet: AH136B tumour (rat hepatoma) cells were implanted subcutaneously (s.c.) in a dorsal site on the foot of male Donryu rats weighing 160–180 g (SLC, Inc., Shizuoka, Japan) with an inoculum size of 1 × 10 7 cells per injection site as described previously ( Doi et al , 1996 ).

Techniques: Injection

Journal: British Journal of Cancer

Article Title: Antiapoptotic effect of haem oxygenase-1 induced by nitric oxide in experimental solid tumour

doi: 10.1038/sj.bjc.6600830

Figure Lengend Snippet: Western blot analysis of HSP70 and HO-1 proteins in AH136B cells and solid tumours. ( A ) Cells were incubated with SNAP (10 or 100 μ M ), P-NONOate (10 or 100 μ M ), or ZnPP IX (100 μ M ) for 6 h or, as a positive control, were treated by heat at 42°C for 30 min, followed by incubation at 37°C for 6 h. ( B ) AH136B solid tumours were treated with 0.9% NaCl solution (control), ZnPP IX (500 μ g kg −1 i.a.), L -NAME (6 mg kg −1 day −1 for 5 days i.p.), or SMT (6 mg kg −1 day −1 for 5 days i.p.). ( C ) Solid tumours were resected at 24 or 48 h after initiation of surgical occlusion of the common iliac artery, which serves the tumour-feeding artery of the tumour-implanted side. Control rats were treated by sham operation. Then, expression of HSP70 and HO-1 proteins was detected by Western blotting by using a monoclonal antibody to mammalian inducible HSP70 or a polyclonal antibody to rat HO-1. Each data point shown represents three independent experiments. See text for details.

Article Snippet: AH136B tumour (rat hepatoma) cells were implanted subcutaneously (s.c.) in a dorsal site on the foot of male Donryu rats weighing 160–180 g (SLC, Inc., Shizuoka, Japan) with an inoculum size of 1 × 10 7 cells per injection site as described previously ( Doi et al , 1996 ).

Techniques: Western Blot, Incubation, Positive Control, Control, Expressing

Journal: Antioxidants

Article Title: Spotlight on Secondary Metabolites Produced by an Early-Flowering Apulian Artichoke Ecotype Sanitized from Virus Infection by Meristem-Tip-Culture and Thermotherapy

doi: 10.3390/antiox13070852

Figure Lengend Snippet: Analysis of rat hepatoma FaO cell response to artichoke extract treatment. ( a ) Integrity of FaO cells exposed to different concentrations of polyphenols resuspended in methanol (MetOH). Extracts were obtained from the old (OL) and young (YL) leaves of BR-NS and BR-S plants. Cytotoxic MetOH effects were evaluated by treating the cells with a scalar dilution of 80% MetOH. ( b ) Dose–response of polyphenol (PP) extracts FaO cells treatment to the accumulation of reactive oxygen species (ROS) in the absence or presence of an oxidative agent (+tBHP). Values represent the means ± standard errors of three independent experiments. CTRL represents the not-treated control used as a reference. (**) double asterisks and (*) single asterisk indicate highly significant and significant differences, respectively, for p ≤ 0.05, in a one-way ANOVA test.

Article Snippet: Briefly, rat hepatoma FaO cells [The European Collection of Authenticated Cell Cultures (EACC)] were plated at a density of 10,000 cells per well in Coon’s modified Ham’s F12 medium supplemented with 10% fetal bovine serum (FBS).

Techniques: Control

Journal: BMC Complementary and Alternative Medicine

Article Title: Isoquercitrin activates the AMP–activated protein kinase (AMPK) signal pathway in rat H4IIE cells

doi: 10.1186/1472-6882-14-42

Figure Lengend Snippet: Effects of isoquercitrin on cell survival and lipid accumulation in H4IIE cells with FFA. (A) H4IIE cells were exposed to various doses of isoquercitrin, and cell viability was measured by MTT tests. Values are expressed as the percentage of the control, which was taken as 100%, and shown as the mean ± SEM (n = 3). (B) and (C) Cells were cultured in the absence or presence of isoquercitrin (50, 100, and 200 μM) for 24 h, followed by stimulation with or without FFA (0.1 mM) for another 24 h. (B) Intracellular oil droplets were visualized by Oil Red O staining under a microscope at 200 × original magnification. (C) Quantitative analysis of lipid deposition was measured by optical density (OD) values at 540 nm after staining. Values are presented as a dose-dependent decrease and expressed as mean ± SEM (n = 3). ** P < 0.01 compared to FFA-treated control group.

Article Snippet: Rat hepatoma (H4IIE) cells, which were purchased from The Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan), were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% streptomycin/penicillin (Nacalai Tesque, Kyoto, Japan) at 37°C in a 5% CO 2 humidified atmosphere.

Techniques: Control, Cell Culture, Staining, Microscopy

Journal: BMC Complementary and Alternative Medicine

Article Title: Isoquercitrin activates the AMP–activated protein kinase (AMPK) signal pathway in rat H4IIE cells

doi: 10.1186/1472-6882-14-42

Figure Lengend Snippet: Isoquercitrin stimulates phosphorylation of AMPK and ACC in H4IIE cells. Cells were incubated with various concentrations of isoquercitrin or control for 12 h. (A) Total cell lysates were subjected to Western blot analysis with antibodies for phosphorylated AMPKα and ACC, total AMPKα, and ACC in H4IIE cells. (B) and (C) Values are shown as mean ± SEM (n = 3). * P < 0.05; ** P < 0.01 compared to control group.

Article Snippet: Rat hepatoma (H4IIE) cells, which were purchased from The Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan), were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% streptomycin/penicillin (Nacalai Tesque, Kyoto, Japan) at 37°C in a 5% CO 2 humidified atmosphere.

Techniques: Phospho-proteomics, Incubation, Control, Western Blot

Journal: BMC Complementary and Alternative Medicine

Article Title: Isoquercitrin activates the AMP–activated protein kinase (AMPK) signal pathway in rat H4IIE cells

doi: 10.1186/1472-6882-14-42

Figure Lengend Snippet: Isoquercitrin decreases the mRNA expression levels of SREBP-1 and FAS in H4IIE cells. Cells were treated with the indicated concentrations of isoquercitrin for 12 h. The mRNA expression levels of SREBP-1 (A) and FAS (B) were assessed by quantitative real-time PCR. Results are expressed as means ± SEM (n = 3) normalized to β-actin mRNA expression. * P < 0.05; ** P < 0.01 compared to control group.

Article Snippet: Rat hepatoma (H4IIE) cells, which were purchased from The Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan), were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% streptomycin/penicillin (Nacalai Tesque, Kyoto, Japan) at 37°C in a 5% CO 2 humidified atmosphere.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control

Journal: BMC Complementary and Alternative Medicine

Article Title: Isoquercitrin activates the AMP–activated protein kinase (AMPK) signal pathway in rat H4IIE cells

doi: 10.1186/1472-6882-14-42

Figure Lengend Snippet: AMPK inhibitor attenuates isoquercitrin-mediated FAS mRNA expression in H4IIE cells. H4IIE cells were incubated with isoquercitrin (200 μM) in the absence or presence of compound C (10 μM) pretreatment for 1 h. (A) Isoquercitrin-induced AMPK activation was suppressed by compound C. (B) The mRNA expression level of FAS was analyzed as indicated. Values are expressed as mean ± SEM (n = 3). ** P < 0.01 between control and isoquercitrin without compound C, ## P < 0.01 between isoquercitrin with or without compound C.

Article Snippet: Rat hepatoma (H4IIE) cells, which were purchased from The Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan), were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% streptomycin/penicillin (Nacalai Tesque, Kyoto, Japan) at 37°C in a 5% CO 2 humidified atmosphere.

Techniques: Expressing, Incubation, Activation Assay, Control

Journal: BMC Complementary and Alternative Medicine

Article Title: Isoquercitrin activates the AMP–activated protein kinase (AMPK) signal pathway in rat H4IIE cells

doi: 10.1186/1472-6882-14-42

Figure Lengend Snippet: Isoquercitrin increases AdipoR1 mRNA expression levels via AMPK in H4IIE cells. (A) Cells were treated with different concentrations of isoquercitrin as mentioned. (B) H4IIE cells were treated with AICAR (0.5 mM and 1 mM) for 12 h as indicated. (C) H4IIE cells were treated with isoquercitrin (200 μM) or AICAR (1 mM) for 12 h in the absence or presence of preincubation with 10 μM compound C for 1 h. The mRNA expression levels of AdipoR1 were measured by quantitative real-time PCR and normalized to β-actin mRNA expression. Values are shown as means ± SEM (n = 3). (A) * P < 0.05; ** P < 0.01 compared to control group. (B) * P < 0.05; ** P < 0.01 compared to control group. (C) ** P < 0.01 between isoquercitrin and control group, ## P < 0.01 between AICAR and control group, N.S. between isoquercitrin and isoquercitrin plus compound C.

Article Snippet: Rat hepatoma (H4IIE) cells, which were purchased from The Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan), were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% streptomycin/penicillin (Nacalai Tesque, Kyoto, Japan) at 37°C in a 5% CO 2 humidified atmosphere.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control

Journal: BMC Complementary and Alternative Medicine

Article Title: Isoquercitrin activates the AMP–activated protein kinase (AMPK) signal pathway in rat H4IIE cells

doi: 10.1186/1472-6882-14-42

Figure Lengend Snippet: AdipoR1 knockdown does not change isoquercitrin-induced AMPK signaling in H4IIE cells. H4IIE cells were transfected with AdipoR1 siRNA or control siRNA as indicated, followed by isoquercitrin treatment for 12 h. (A) After transfection, AdipoR1 expression was determined by quantitative RT-PCR. (B) Phosphorylation and total AMPKα protein expression were measured by Western blotting. Results were analyzed and shown as means ± SEM (n = 3). # P < 0.05 between isoquercitrin and control plus AdipoR1 siRNA.

Article Snippet: Rat hepatoma (H4IIE) cells, which were purchased from The Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan), were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% streptomycin/penicillin (Nacalai Tesque, Kyoto, Japan) at 37°C in a 5% CO 2 humidified atmosphere.

Techniques: Knockdown, Transfection, Control, Expressing, Quantitative RT-PCR, Phospho-proteomics, Western Blot

Journal: The Journal of Cell Biology

Article Title: Inhibitors of Copi and Copii Do Not Block PEX3 -Mediated Peroxisome Synthesis

doi:

Figure Lengend Snippet: PEX3 encodes a peroxisomal protein. A postnuclear supernatant of HepG2 cells was fractionated by Nycodenz gradient centrifugation. Equal proportions of each fraction were assayed for the marker enzymes catalase (a peroxisomal marker), SDH (a mitochondrial marker), and NCR (a microsomal marker). Equal proportions of each fraction were also assayed by immunoblot with antibodies to PEX3. A 42-kD band corresponding to PEX3 was detected exclusively in the peroxisomal fractions.

Article Snippet: HepG2 cells were obtained from Michael Schrader (The Johns Hopkins University, Baltimore, MD).

Techniques: Gradient Centrifugation, Marker, Western Blot