Journal: Oncology reports

Article Title: Notch1 is a potential therapeutic target for the treatment of human hepatitis B virus X protein-associated hepatocellular carcinoma.

doi: 10.3892/or.2013.2917

Figure Lengend Snippet: Figure 1. Notch1-shRNA suppresses L02/HBx cell proliferation in vitro. (A) Identification of the effective shRNA targeting the Notch1 gene. The relative mRNA and protein levels of Notch1 were assessed by qRT-PCR and western blotting in L02/HBx cells 48 h after transient transfection with Notch1-shRNA1, Notch1-shRNA2 or negative control-shRNA (NC), respectively. (B) The components of the Notch1 signaling pathway were downregulated in the L02/ HBx-Notch1 shRNA2 cells. The mRNA and protein expression levels of Notch1 and Hes1 were assessed by qRT-PCR and western blotting, respectively. Actin was used as a loading control for both quantitative RT-PCR and western blotting. (C) CCK-8 assay and (D) colony formation assay of L02/HBx cells stably transfected with control or Notch1 shRNA2. Data are shown as the mean ± SEM from at least three independent experiments. Statistically significant differences are indicated as *P<0.05, **P<0.01 vs. L02/HBx cells.

Article Snippet: For immunohistochemical analysis, staining for Notch1 was carried out using rabbit polyclonal antibodies against Notch1 (Proteintech Group) at a dilution of 1:50.

Techniques: shRNA, In Vitro, Quantitative RT-PCR, Western Blot, Transfection, Negative Control, Expressing, Control, CCK-8 Assay, Colony Assay, Stable Transfection

Journal: Oncology reports

Article Title: Notch1 is a potential therapeutic target for the treatment of human hepatitis B virus X protein-associated hepatocellular carcinoma.

doi: 10.3892/or.2013.2917

Figure Lengend Snippet: Figure 2. Notch1-shRNAs suppresses L02/HBx cell proliferation in vivo. (A) The growth curve of the tumors derived from L02/HBx cells pretreated with Notch1-shRNA or control-shRNA in nude mice. Data represent means ± SEM of six samples. *P<0.05, **P<0.01. (B) Images of representative mice and dis sected tumors from nude mice. (C) Tumor tissues from nude mice inoculated with NC or Notch1-shRNA cells were stained with hematoxylin and eosin (H&E). Original magnification, x200. (D) Immunohistochemistry of Notch1 expression in tumor tissues from nude mice. Original magnification, x400.

Article Snippet: For immunohistochemical analysis, staining for Notch1 was carried out using rabbit polyclonal antibodies against Notch1 (Proteintech Group) at a dilution of 1:50.

Techniques: In Vivo, Derivative Assay, shRNA, Control, Staining, Immunohistochemistry, Expressing

Journal: Oncology reports

Article Title: Notch1 is a potential therapeutic target for the treatment of human hepatitis B virus X protein-associated hepatocellular carcinoma.

doi: 10.3892/or.2013.2917

Figure Lengend Snippet: Figure 3. Notch1-shRNA induces cell cycle arrest via the CyclinD1/CDK4 pathway in L02/HBx cells. (A) Notch1-shRNA induced cell cycle arrest in L02/ HBx cells. Cell cycle distribution was examined by flow cytometry after staining with PI. Results are visualized as a representative experiment or means ± SEM of three experiments. (B) Notch1-shRNA downregulates the CyclinD1/CDK4 pathway in L02/HBx cells. The expression levels of cell cycle regulatory genes CyclinD1, CDK4, E2F1, Rb, p21 and CyclinE1 were analyzed by qRT-PCR and western blotting. Data represent the mean ± SEM from three independent experiments. Statistically significant differences are indicated as *P<0.05, **P<0.01 vs. L02/HBx cells.

Article Snippet: For immunohistochemical analysis, staining for Notch1 was carried out using rabbit polyclonal antibodies against Notch1 (Proteintech Group) at a dilution of 1:50.

Techniques: shRNA, Cytometry, Staining, Expressing, Quantitative RT-PCR, Western Blot

Journal: Oncology reports

Article Title: Notch1 is a potential therapeutic target for the treatment of human hepatitis B virus X protein-associated hepatocellular carcinoma.

doi: 10.3892/or.2013.2917

Figure Lengend Snippet: Figure 4. Notch1-shRNA increases cell apoptosis via the caspase-9-caspase-3 pathway in L02/HBx cells. (A) Notch1-shRNA increased cell apoptosis in L02/ HBx cells. Analysis of apoptosis was performed using PE Annexin V and 7-AAD staining by FACS analysis. Results are visualized as a representative experi ment or means ± SEM of three experiments. (B) Notch1-shRNA upregulated the caspase-9-caspase-3 pathway in L02/HBx cells. The expression of caspase-9, -8 and -3 was analyzed by qRT-PCR and western blotting. Data are shown as the mean ± SEM of three independent experiments. Statistically significant differences are indicated as *P<0.05, **P<0.01 vs. L02/HBx cells.

Article Snippet: For immunohistochemical analysis, staining for Notch1 was carried out using rabbit polyclonal antibodies against Notch1 (Proteintech Group) at a dilution of 1:50.

Techniques: shRNA, Staining, Expressing, Quantitative RT-PCR, Western Blot

Journal: Oncogene

Article Title: Lunatic Fringe is a potent tumor suppressor in Kras-initiated pancreatic cancer.

doi: 10.1038/onc.2015.306

Figure Lengend Snippet: Figure 1. Lfng is expressed in a subset of acinar cells in the adult pancreas. (a–f) Expression of Lfng in the adult pancreas. Shown are representative photomicrographs of whole-mount (a–c) and sectioned. (d–f) X-Gal staining of the pancreas from wild-type, LfnglacZ/+ and LfnglacZ/lacZ mice at 6 months of age. (g–j) Expression of individual Notch receptors in the adult pancreas. Shown are representative photomicrographs of anti-Notch1, anti-Notch2, anti-Notch3 and anti-Notch4 immunostaining in the wild pancreas at 6 months of age. Arrow in g inset points to Notch1 expression in the blood vessel. Arrows in i and j indicate Notch3 expression in ductal cells and Notch4 expression in islet, respectively. Scale bars: 50 μm.

Article Snippet: Primary antibodies used for immunostaining were: Notch1 (Cell Signaling, Danvers, MA, USA; No. 3608, 1:100), Notch2 (DSHB, University of Iowa, C651.6DbHN, 1:200), Notch3 (ProteinTech, Chicago, IL, USA; 55114-1-AP, 1:100), Notch4 (Millipore, Billerica, MA, USA; 09-089, 1:100), Ki67 (Abcam, Cambridge, MA, USA; ab16667, 1:100), Aldh1 (Abcam, ab52492, 1:200), CK19 (DSHB, University of Iowa, TROMA-III, 1:400), E-cadherin (Cell Signaling, No. 3195, 1:100), Vimentin (Cell Signaling, No. 5741, 1:100), Clusterin (Santa Cruz, Dallas, TX, USA; sc-6420, 1:50), Phospho-Smad2 (Ser465/467) (Cell Signaling, No. 3101, 1:100), and GFP (Invitrogen, Grand Island, NY, USA; A11122, 1:200).

Techniques: Expressing, Staining, Immunostaining

Journal: Oncogene

Article Title: Lunatic Fringe is a potent tumor suppressor in Kras-initiated pancreatic cancer.

doi: 10.1038/onc.2015.306

Figure Lengend Snippet: Figure 2. Deletion of Lfng enhanced Notch signaling in the KrasLSL-G12D/+;Pdx1-Cre pancreas. (a) Quantitative RT–PCR for Lfng and Notch downstream target gene Hes1 in the pancreas from Pdx1-Cre (C), Lfngflox/flox;Pdx1-Cre (LC), KrasLSL-G12D/+;Pdx1-Cre (KC) and Lfngflox/flox;KrasLSL-G12D/+; Pdx1-Cre (LKC) mice of indicated age. *Po0.05; **Po0.005; ***Po0.0005. (b) Western blot analysis of individual Notch receptors in the pancreas from C and KC mice at 3 months of age, with quantification of band densities relative to β-actin. (c) Western blot analysis of Notch receptors in the pancreas from KC and LKC mice of indicated age, and quantification of band densities relative to β-actin. (d) Representative photomicrographs of anti-Notch1 and anti-Notch3 immunostaining in the pancreas from 3 and 5-month-old KC and LKC mice. Arrows point to ADM lesions in 3-month-old mice and ductal cells in 5-month-old mice, respectively. Scale bars: 50 μm.

Article Snippet: Primary antibodies used for immunostaining were: Notch1 (Cell Signaling, Danvers, MA, USA; No. 3608, 1:100), Notch2 (DSHB, University of Iowa, C651.6DbHN, 1:200), Notch3 (ProteinTech, Chicago, IL, USA; 55114-1-AP, 1:100), Notch4 (Millipore, Billerica, MA, USA; 09-089, 1:100), Ki67 (Abcam, Cambridge, MA, USA; ab16667, 1:100), Aldh1 (Abcam, ab52492, 1:200), CK19 (DSHB, University of Iowa, TROMA-III, 1:400), E-cadherin (Cell Signaling, No. 3195, 1:100), Vimentin (Cell Signaling, No. 5741, 1:100), Clusterin (Santa Cruz, Dallas, TX, USA; sc-6420, 1:50), Phospho-Smad2 (Ser465/467) (Cell Signaling, No. 3101, 1:100), and GFP (Invitrogen, Grand Island, NY, USA; A11122, 1:200).

Techniques: Quantitative RT-PCR, Western Blot, Immunostaining

Journal: Food Science & Nutrition

Article Title: Vitamin D ameliorates asthma‐induced lung injury by regulating HIF‐1α/Notch1 signaling during autophagy

doi: 10.1002/fsn3.2880

Figure Lengend Snippet: The primer sequences

Article Snippet: G1285 and G1243, respectively; Solarbio Life Science); interleukin (IL)‐6, IL‐10, and IL‐17 enzyme‐linked immunoassay (ELISA) kits; (Shanghai Jingtian Biotechnology Co., Ltd.); TUNEL kit (cat. no. KGA703; KeyGen Biotech); rabbit antimouse primary antibody LC 3B (cat. no. ab225383; Abcam), Beclin1 (cat. no. ab207612; Abcam), P62 (cat. no. ab91526; Abcam), HIF‐1α (cat. no. ab51608; Abcam), and Notch1 (cat. no. ab276343; Abcam); GAPDH (cat. no. ab181602; Abcam); goat antirabbit secondary antibody (batch No. SA00001‐2; Proteintech); Trizol reagent (batch No. 15596026; Ambion, Inc.); reverse transcription kit (batch no.: K1622; Thermo Scientific); real‐time fluorescent quantitative polymerase chain reaction (real‐time qPCR) kit (batch No. PA0025; Shanghai Jinan Biotechnology Co., Ltd.).

Techniques:

Journal: Food Science & Nutrition

Article Title: Vitamin D ameliorates asthma‐induced lung injury by regulating HIF‐1α/Notch1 signaling during autophagy

doi: 10.1002/fsn3.2880

Figure Lengend Snippet: HIF‐1α (hypoxia‐inducible factor 1α) and Notch1 (neurogenic locus notch homolog protein 1) expression by western blotting analysis. Normal: Mice treated with normal saline; Model: asthmatic model mice treated with normal saline; LD: asthmatic model mice treated with low‐dose Vit D (100 IU/kg/day); MD: asthmatic model mice treated with middle‐dose Vit D (500 IU/kg/day); HD: asthmatic model mice treated with high‐dose Vit D (1000 IU/kg/day). *** p < .001, compared with Normal; # p < .05, ## p < .01, ### p < .001, compared with Model; $ p < .05, $$ p < .01, compared with LD group; & p < .05, compared with MD group

Article Snippet: G1285 and G1243, respectively; Solarbio Life Science); interleukin (IL)‐6, IL‐10, and IL‐17 enzyme‐linked immunoassay (ELISA) kits; (Shanghai Jingtian Biotechnology Co., Ltd.); TUNEL kit (cat. no. KGA703; KeyGen Biotech); rabbit antimouse primary antibody LC 3B (cat. no. ab225383; Abcam), Beclin1 (cat. no. ab207612; Abcam), P62 (cat. no. ab91526; Abcam), HIF‐1α (cat. no. ab51608; Abcam), and Notch1 (cat. no. ab276343; Abcam); GAPDH (cat. no. ab181602; Abcam); goat antirabbit secondary antibody (batch No. SA00001‐2; Proteintech); Trizol reagent (batch No. 15596026; Ambion, Inc.); reverse transcription kit (batch no.: K1622; Thermo Scientific); real‐time fluorescent quantitative polymerase chain reaction (real‐time qPCR) kit (batch No. PA0025; Shanghai Jinan Biotechnology Co., Ltd.).

Techniques: Expressing, Western Blot