Journal: Molecular Medicine Reports

Article Title: CSN1 facilitates proliferation and migration of hepatocellular carcinoma cells by upregulating cyclin A2 expression

doi: 10.3892/mmr.2020.11684

Figure Lengend Snippet: Relative CSN1 expression in HCC and its clinical significance. (A) Tissue microarray immunohistochemical staining. Magnification, ×200. (B) Relative expression of CSN1 in HCC tissue (n=117) and paracancerous tissue samples (n=59). (C) CSN1 expression levels in cancer and adjacent tissue samples. n=59 in each group. (D and E) Kaplan-Meier curves for (D) overall survival or (E) disease-free survival in two groups defined by low and high expression of CSN1 in patients with HCC. (F) CSN1 protein levels in HCC cell lines compared with the normal human liver cell line MIHA. **P<0.01 and ***P<0.001. CSN1, constitutive photomorphogenesis 9 signalosome subunit 1; HCC, hepatocellular carcinoma.

Article Snippet: The membrane was blocked with 5% non-fat milk in TBS-Tween-20 for 1 h at room temperature, incubated with the indicated primary antibodies overnight at 4°C, and then with horseradish peroxidase-conjugated secondary antibodies (1:5,000; ProteinTech Group, Inc.) at room temperature for 1 h. The primary antibodies used were specific for CSN1 (1:2,000; cat. no. 11709-1-AP; ProteinTech Group, Inc.), cyclin A2 (1:2,000, cat. no. GTX103042; GeneTex, Inc.), CDK-2 (1:2,000; cat. no. 10122-1-AP; ProteinTech Group, Inc.), CDK-4 (1:2,000; cat. no. 11026-1-AP; ProteinTech Group, Inc.), phospho (p)-CDK4 (1:2,000, cat. no. GTX00778; GeneTex, Inc.) or GAPDH (1:5,000; cat. no. 10494-1-AP; ProteinTech Group, Inc.).

Techniques: Expressing, Microarray, Immunohistochemical staining, Staining

Journal: Molecular Medicine Reports

Article Title: CSN1 facilitates proliferation and migration of hepatocellular carcinoma cells by upregulating cyclin A2 expression

doi: 10.3892/mmr.2020.11684

Figure Lengend Snippet: Relationship between the expression of CSN1 and clinicopathological characteristics of patients with hepatocellular carcinoma.

Article Snippet: The membrane was blocked with 5% non-fat milk in TBS-Tween-20 for 1 h at room temperature, incubated with the indicated primary antibodies overnight at 4°C, and then with horseradish peroxidase-conjugated secondary antibodies (1:5,000; ProteinTech Group, Inc.) at room temperature for 1 h. The primary antibodies used were specific for CSN1 (1:2,000; cat. no. 11709-1-AP; ProteinTech Group, Inc.), cyclin A2 (1:2,000, cat. no. GTX103042; GeneTex, Inc.), CDK-2 (1:2,000; cat. no. 10122-1-AP; ProteinTech Group, Inc.), CDK-4 (1:2,000; cat. no. 11026-1-AP; ProteinTech Group, Inc.), phospho (p)-CDK4 (1:2,000, cat. no. GTX00778; GeneTex, Inc.) or GAPDH (1:5,000; cat. no. 10494-1-AP; ProteinTech Group, Inc.).

Techniques: Expressing

Journal: Molecular Medicine Reports

Article Title: CSN1 facilitates proliferation and migration of hepatocellular carcinoma cells by upregulating cyclin A2 expression

doi: 10.3892/mmr.2020.11684

Figure Lengend Snippet: Effects of CSN1 on hepatocellular carcinoma cell proliferation, cell cycle and apoptosis. (A) CSN1 expression levels in MHCC-LM3, Hep3B and MHCC-97H cells transfected with si-CSN1-1, −2 and −3 or siNC. (B) Proliferation of MHCC-LM3 and Hep3B cells following CSN1 knockdown. Magnification, ×200. (C) Representative images (upper panel) and quantification (lower panel) of the Transwell assays for MHCC-LM3 and Hep3B cells transfected with si-CSN1 or si-NC. Magnifications, ×100 and ×400. (D) Measurement of the migration rate after CSN1 knockdown in MHCC-LM3 and Hep3B cells. Magnification, ×100. (E) Apoptosis and (F) cell cycle measurement following CSN1 silencing in MHCC-LM3 and Hep3B cells. *P<0.05, **P<0.01 and ***P<0.001. CSN1, constitutive photomorphogenesis 9 signalosome subunit 1; si, small interfering RNA; NC, negative control.

Article Snippet: The membrane was blocked with 5% non-fat milk in TBS-Tween-20 for 1 h at room temperature, incubated with the indicated primary antibodies overnight at 4°C, and then with horseradish peroxidase-conjugated secondary antibodies (1:5,000; ProteinTech Group, Inc.) at room temperature for 1 h. The primary antibodies used were specific for CSN1 (1:2,000; cat. no. 11709-1-AP; ProteinTech Group, Inc.), cyclin A2 (1:2,000, cat. no. GTX103042; GeneTex, Inc.), CDK-2 (1:2,000; cat. no. 10122-1-AP; ProteinTech Group, Inc.), CDK-4 (1:2,000; cat. no. 11026-1-AP; ProteinTech Group, Inc.), phospho (p)-CDK4 (1:2,000, cat. no. GTX00778; GeneTex, Inc.) or GAPDH (1:5,000; cat. no. 10494-1-AP; ProteinTech Group, Inc.).

Techniques: Expressing, Transfection, Knockdown, Migration, Small Interfering RNA, Negative Control

Journal: Molecular Medicine Reports

Article Title: CSN1 facilitates proliferation and migration of hepatocellular carcinoma cells by upregulating cyclin A2 expression

doi: 10.3892/mmr.2020.11684

Figure Lengend Snippet: CSN1 interacts with cyclin A2. (A) Representative western blots of CSN1, cyclin A2 and cell cycle-related proteins following CSN1 overexpression in MHCC-LM3 cells. Cells transfected with empty vectors were used as NC group. (B) Western blots of cyclin A2 protein levels in MHCC-LM3 cells transfected with NC or OE-CSN1 plasmids, with or without si-cyclin A2. (C) Proliferation (magnification, ×200), (D) wound healing (magnification, ×100) (E) and Transwell assay (magnification, ×100 and ×400) of MHCC-LM3 cells transfected with NC or OE-CSN1 plasmids, with or without si-cyclin A2. (F) Effect of OE-CSN1 on cyclin A2 mRNA levels. (G) Representative western blots of cyclin A2 turnover rate in OE-CSN1 MHCC-LM3 cells following CXH treatment. (H) Cyclin A2 protein levels after OE-CSN1 transfection, and CHX or ActD treatment in MHCC-LM3 cells. *P<0.05, **P<0.01 and ***P<0.001. CHX, cycloheximide; CSN1, constitutive photomorphogenesis 9 signalosome subunit 1; OE, overexpression; ActD, actinomycin D; CDK, cyclin-dependent kinase; si, small interfering; NC, negative control.

Article Snippet: The membrane was blocked with 5% non-fat milk in TBS-Tween-20 for 1 h at room temperature, incubated with the indicated primary antibodies overnight at 4°C, and then with horseradish peroxidase-conjugated secondary antibodies (1:5,000; ProteinTech Group, Inc.) at room temperature for 1 h. The primary antibodies used were specific for CSN1 (1:2,000; cat. no. 11709-1-AP; ProteinTech Group, Inc.), cyclin A2 (1:2,000, cat. no. GTX103042; GeneTex, Inc.), CDK-2 (1:2,000; cat. no. 10122-1-AP; ProteinTech Group, Inc.), CDK-4 (1:2,000; cat. no. 11026-1-AP; ProteinTech Group, Inc.), phospho (p)-CDK4 (1:2,000, cat. no. GTX00778; GeneTex, Inc.) or GAPDH (1:5,000; cat. no. 10494-1-AP; ProteinTech Group, Inc.).

Techniques: Western Blot, Over Expression, Transfection, Transwell Assay, Negative Control

Journal: Molecular Medicine Reports

Article Title: CSN1 facilitates proliferation and migration of hepatocellular carcinoma cells by upregulating cyclin A2 expression

doi: 10.3892/mmr.2020.11684

Figure Lengend Snippet: Effects of CSN1 on tumor growth in vivo. Nude mice were subcutaneously injected with CSN1-overexpressing MHCC-LM3 cells and NC cells. Tumor sizes were measured after sizeable tumor formation (day 8). On day 28, the tumors were excised for analysis. n=5. (A) Western blot images of CSN1 levels. (B) Macroscopic appearance of isolated tumors. (C) Tumor volume and (D) weights in CSN1 and NC xenograft nude mice. *P<0.05 and **P<0.01. CSN1, constitutive photomorphogenesis 9 signalosome subunit 1; NC, negative control.

Article Snippet: The membrane was blocked with 5% non-fat milk in TBS-Tween-20 for 1 h at room temperature, incubated with the indicated primary antibodies overnight at 4°C, and then with horseradish peroxidase-conjugated secondary antibodies (1:5,000; ProteinTech Group, Inc.) at room temperature for 1 h. The primary antibodies used were specific for CSN1 (1:2,000; cat. no. 11709-1-AP; ProteinTech Group, Inc.), cyclin A2 (1:2,000, cat. no. GTX103042; GeneTex, Inc.), CDK-2 (1:2,000; cat. no. 10122-1-AP; ProteinTech Group, Inc.), CDK-4 (1:2,000; cat. no. 11026-1-AP; ProteinTech Group, Inc.), phospho (p)-CDK4 (1:2,000, cat. no. GTX00778; GeneTex, Inc.) or GAPDH (1:5,000; cat. no. 10494-1-AP; ProteinTech Group, Inc.).

Techniques: In Vivo, Injection, Western Blot, Isolation, Negative Control

Journal: Journal of the Royal Society Interface

Article Title: Ontogenetic development of migration: Lagrangian drift trajectories suggest a new paradigm for sea turtles

doi: 10.1098/rsif.2010.0009

Figure Lengend Snippet: Attachment details for tracked adult turtles. Data for turtles A–G are from Zbinden et al . (2008) . Turtles 4394 and 15119_07 are described in Schofield et al . (in press) . Argos quality locations were provided from the KiwiSat 101 and SMRU solar satellite tags. The Sirtrack GPS-Argos and SMRU GPS-GSM tags provided Fastloc GPS locations. The GPS-GSM tag relayed data via mobile phone receivers. All the other tags relayed data via the Argos satellite system. General foraging location indicated with ‘Greece’ means those that had foraging locations in Greece excluding Zakynthos.

Article Snippet: The Sirtrack GPS-Argos and SMRU GPS-GSM tags provided Fastloc GPS locations.

Techniques: