Journal: Nutrients

Article Title: Walnut Phenolic Extract and Its Bioactive Compounds Suppress Colon Cancer Cell Growth by Regulating Colon Cancer Stemness

doi: 10.3390/nu8070439

Figure Lengend Snippet: Primers for reverse transcriptase polymerase chain reaction RT-PCR.

Article Snippet: The membranes were subsequently blocked in 5% non-fat dried milk in Tris-buffered saline (TBS) and were incubated with primary antibodies against cytokeratin 20 (CK20, Abcam, Cambridge, MA, USA), Notch1 (Novus Biologicals, Littletown, CO, USA), β-catenin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and p-GSK3β (Cell Signaling Technology, Danvers, MA, USA) overnight.

Techniques: Reverse Transcription, Polymerase Chain Reaction

Journal: Nutrients

Article Title: Walnut Phenolic Extract and Its Bioactive Compounds Suppress Colon Cancer Cell Growth by Regulating Colon Cancer Stemness

doi: 10.3390/nu8070439

Figure Lengend Snippet: WPE and its bioactive compounds suppress colon CSCs markers, including CD133, CD44, DLK1, and Notch1 as well as Wnt/β-catenin signaling in colon CSCs. CD133 + CD44 + HCT116 cells were treated with varying concentrations of WPE (0, 10, 20, and 40 μg/mL) ( A a , Ba ); or concentrations of (+)-catechin, chlorogenic acid, ellagic acid and gallic acid comparable to 40 μg/mL of WPE ( A b , Bb ) for 6 days. mRNA expressions of CD133, CD44, DLK1 and Notch1 were examined by RT-PCR, and β-actin was used as a loading control ( A ); Protein levels of Notch1, β-catenin and p-GSK3β were examined by Western blot analysis, and α-tubulin was used as a loading control ( B ). Representative blots are shown in left panel and quantified in right panel. The values shown are the means ± SEM. A p -value < 0.05 was considered significant. Ctrl, Control; WPE, walnut phenolic extract; CSCs, cancer stem cells; CAT, (+)-catechin; CGA, chlorogenic acid; EA, ellagic acid; GA, gallic acid.

Article Snippet: The membranes were subsequently blocked in 5% non-fat dried milk in Tris-buffered saline (TBS) and were incubated with primary antibodies against cytokeratin 20 (CK20, Abcam, Cambridge, MA, USA), Notch1 (Novus Biologicals, Littletown, CO, USA), β-catenin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and p-GSK3β (Cell Signaling Technology, Danvers, MA, USA) overnight.

Techniques: Reverse Transcription Polymerase Chain Reaction, Control, Western Blot

Journal: Nutrients

Article Title: Walnut Phenolic Extract and Its Bioactive Compounds Suppress Colon Cancer Cell Growth by Regulating Colon Cancer Stemness

doi: 10.3390/nu8070439

Figure Lengend Snippet: WPE down-regulates CD133, CD44, DLK1, and Notch1 in human primary cells from colorectal cancer tissue. Primary cancer cells were treated with varying concentrations of WPE (0, 10, 20, and 40 μg/mL) for 3 d. mRNA expressions of CD133, CD44, DLK1 and Notch1 were detected by RT-PCR, and GAPDH was used as a loading control. The values shown are the means ± SEM. A p -value < 0.05 was considered significant. Ctrl, Control.

Article Snippet: The membranes were subsequently blocked in 5% non-fat dried milk in Tris-buffered saline (TBS) and were incubated with primary antibodies against cytokeratin 20 (CK20, Abcam, Cambridge, MA, USA), Notch1 (Novus Biologicals, Littletown, CO, USA), β-catenin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and p-GSK3β (Cell Signaling Technology, Danvers, MA, USA) overnight.

Techniques: Reverse Transcription Polymerase Chain Reaction, Control