Figure 3

Effects on the cell cycle, and phosphorylated GSK-3β, GSK-3β, β-catenin, and cyclin D1 in H9c2 cells. (a) Top: Cell cycle phase determined by flow cytometry for H9c2 cells in the control medium (left top, label C), or in the medium containing 20 μM EGCg for 30 min (right top, label E), or in the medium containing 400 μM H₂O₂ for 30 min with (right bottom, label E + H) or without 20 μM EGCg pretreatment for 30 min (left bottom, label H). Bottom: Quantitative analysis of the cell cycle phase. (b) Western blotting (top) with quantitative analyses (bottom) of pGSK-3β, GSK-3β, and GAPDH levels for the H9c2 cells cultured in the medium as indicated. (c) Western blotting (top) with quantitative analyses (bottom) of β-catenin, cyclin D1, and β-actin levels for the H9c2 cells in the medium with the addition as indicated. (d) Western blots showing inhibition of GSK-3β on β-catenin levels in the H9c2 cells. Lane 1: cells cultured in the control medium; lane 2: cells cultured in the medium containing 20 μM EGCg for 30 min; lane 3: cells cultured in the medium containing 400 μM H₂O₂ for 30 min, lane 4: cells cultured in the medium containing 400 μM H₂O₂ for 30 min with 20 μM EGCg pre-treatment for 30 min, lane 5: cells cultured in the medium containing 400 μM H₂O₂ for 30 min with the pretreatment of 10 μM SB 216763 inhibitor of GSK-3α/3β for 30 min. In a, b, and c, the values are the mean ± SEM (n = 6), with *, # indicating a significant difference compared to the cells in control medium and the cells treated with H₂O₂, respectively.