Journal: Oncology Letters

Article Title: Potential use of chymotrypsin-like proteasomal activity as a biomarker for prostate cancer

doi: 10.3892/ol.2018.7936

Figure Lengend Snippet: Proteasomal activity in vitro. (A) Chymotrypsin-like, caspase-like and trypsin-like activity of the proteasome was measured in LNCaP and prostate epithelial cells. Chymotrypsin-like and caspase-like activities were significantly different between the LNCaP and prostate epithelial cells. Data are presented as means ± standard deviation. *P<0.01. (B) Representative images of western blotting to determine the expression of Bax, IκB-α and p27 in LNCaP cells compared with prostate epithelial cells, with GAPDH as a loading control. (C) Quantified western blotting results. Each bar represents the mean of three independent experiments; data are expressed as mean ± standard deviation (SD). *P<0.01. Bax, Bcl-2-associated X; IκB-α, NF-κB inhibitor-α.

Article Snippet: Mouse monoclonal antibodies against Bax (cat. no. sc-23959) and p27 (cat. no. sc-1641), and rabbit polyclonal antibodies against inhibitor of nuclear factor (NF)-κB-α (IκB-α; cat. no. sc-203) and GAPDH (FL-335; cat. no. sc-25778) were both from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

Techniques: Activity Assay, In Vitro, Standard Deviation, Western Blot, Expressing, Control

Journal: Oncology Letters

Article Title: Potential use of chymotrypsin-like proteasomal activity as a biomarker for prostate cancer

doi: 10.3892/ol.2018.7936

Figure Lengend Snippet: Expression of Bax, IκB-α, and p27 protein in vivo. (A) Representative images of western blotting to determine the expression of Bax, IκB-α and p27 in the mouse tumor tissue and the prostate tissue from the control mice. Tumor tissue was removed on day 29 for analysis by western blotting. Decreased Bax, IκBa, and p27 protein levels were observed in the tumor tissues, with GAPDH as a loading control. (B) Quantified western blotting results. Three mice were selected from each group for this assay. Each bar represents the mean of three independent experiments; data were expressed as the mean ± standard deviation (SD). *P<0.01. Bax, Bcl-2-associated X; IκB-α, NF-κB inhibitor-α.

Article Snippet: Mouse monoclonal antibodies against Bax (cat. no. sc-23959) and p27 (cat. no. sc-1641), and rabbit polyclonal antibodies against inhibitor of nuclear factor (NF)-κB-α (IκB-α; cat. no. sc-203) and GAPDH (FL-335; cat. no. sc-25778) were both from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

Techniques: Expressing, In Vivo, Western Blot, Control, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Nucleoside 5′-Phosphoramidates Control the Phenylpropanoid Pathway in Vitis vinifera Suspension-Cultured Cells

doi: 10.3390/ijms222413567

Figure Lengend Snippet: Expression of stilbene synthase gene ( STS1) and accumulation of stilbenes in cells of Vitis vinifera cv. Monastrell treated with 5 µM nucleoside 5′-phosphoramidates. ( a ) STS1 , stilbene synthase; ( b ) accumulation of trans -resveratrol; ( c ) accumulation of trans -piceid. Total RNA was reverse-transcribed into cDNA and used as a template for real-time quantification PCR reaction as described in the . Specific primers were designed for STS1 and EFα1 (elongation factor 1-alpha, which was used as an endogenous control). The expression level of STS1 in the control cells (no nucleotide added) was set to 1. Values represent the mean ± standard deviation of the three replicates. Accumulation of trans -resveratrol and trans -piceid was determined using the HPLC method as described in the . Values without a common letter are significantly different according to ANOVA and Tukey’s HSD multiple range test ( p ≤ 0.05).

Article Snippet: To determine the intracellular content of trans -resveratrol and trans -piceid, 200 mg of freeze-dried cells were extracted overnight with 4 mL of methanol at 4 °C with continuous shaking, and then 20 μL of each sample was analyzed on a LiChrospher 100 RP-18 column as described above. trans -Resveratrol and trans -piceid were identified (at 308 nm) and quantified by comparison with standards of commercial trans -resveratrol (Sigma-Aldrich, St. Louis, MO, USA) and trans -piceid (ChromaDex, Los Angeles, CA, USA) using respective calibration curves.

Techniques: Expressing, Reverse Transcription, Control, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Nucleoside 5′-Phosphoramidates Control the Phenylpropanoid Pathway in Vitis vinifera Suspension-Cultured Cells

doi: 10.3390/ijms222413567

Figure Lengend Snippet: Expression of the VvABCG44 resveratrol transporter gene in the cells of Vitis vinifera cv. Monastrell treated with 5 µM nucleoside 5′-phosphoramidates, and accumulation of stilbenes in the spent medium. ( a ) VvABCG44 , resveratrol transporter gene; ( b ) accumulation of trans -resveratrol; ( c ) accumulation of trans -piceid. Total RNA was reverse-transcribed into cDNA and used as a template for real-time quantification PCR reaction as described in the . Specific primers were designed for VvABCG44 and EFα1 (elongation factor 1-alpha, which was used as an endogenous control). The expression level of VvABCG44 in the control cells (no nucleotide added) was set to 1. Accumulation of trans -resveratrol and trans -piceid was determined using the HPLC method as described in the . Values are the mean ± standard deviation of the three replicates. Values without a common letter were significantly different according to ANOVA and Tukey’s HSD multiple range test ( p ≤ 0.05).

Article Snippet: To determine the intracellular content of trans -resveratrol and trans -piceid, 200 mg of freeze-dried cells were extracted overnight with 4 mL of methanol at 4 °C with continuous shaking, and then 20 μL of each sample was analyzed on a LiChrospher 100 RP-18 column as described above. trans -Resveratrol and trans -piceid were identified (at 308 nm) and quantified by comparison with standards of commercial trans -resveratrol (Sigma-Aldrich, St. Louis, MO, USA) and trans -piceid (ChromaDex, Los Angeles, CA, USA) using respective calibration curves.

Techniques: Expressing, Reverse Transcription, Control, Standard Deviation