Journal: Nutrition Research and Practice
Article Title: Ethanol extract of Innotus obliquus (Chaga mushroom) induces G1 cell cycle arrest in HT-29 human colon cancer cells
Figure Lengend Snippet: Effect of EEIO on HT-29 cell growth. HT-29 cells were plated in 24-well plates at a density of 50,000 cells/well in DMEM/F12 supplemented with 10% FBS. One day later, the monolayers were serum-starved with serum-free DMEM/F12 supplemented with 5 mg/L transferrin, 0.1 g/L BSA, and 5 µg/mL selenium for 24 h. After serum starvation, cells were incubated in serum-free medium in the absence or presence of various concentrations of EEIO. Cell numbers were estimated by MTT assay. Each bar represents the mean ± SEM (n = 6). Bars with different letters are significantly different at P
Article Snippet: Materials The reagents used in this study were purchased from the following suppliers: Dulbecco's modified Eagle's medium/Ham's F12 nutrient mixture (DMEM/F12) and selenium from Gibco BRL (Gaithersburg, MD, USA); fetal bovine serum (FBS), trypsin-EDTA, and penicillin/streptomycin from Cambrex Bio Technology (Walkersville, MD, USA); 3-[4,5-dimetjylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT), anti-β-actin, RIA-grade bovine serum albumin (BSA), and transferrin from Sigma-Aldrich Co. (St. Louis, MO, USA); antibodies against cyclin D1 and phospho-Rb (Ser807/811) from Cell Signaling Technology (Beverly, MA, USA); antibodies against p21CIP1/WAF1 (c-19), p27KIP1 , p53, CDK2 (M-2), CDK4 (c-22), E2F-1 (C-20), and Rb (c-15) from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
Techniques: Incubation, MTT Assay