Figure 2
Ergosterol inhibited the β-catenin signaling pathway. (a) The effects of ER treatment (2 and 20 μM) were assessed by conducting Western blot analysis on β-catenin, c-Myc, and cyclin D1 after a 24 h period. (b) The translocation of β-catenin to the nucleus was evaluated using immunofluorescence staining after subjecting cells to 24 h treatments with DMSO or ER 20 at μM. Co-localization of β-catenin and the nucleus was visualized using confocal microscopy at a magnification of 200X, aided by the application of Hoechst 33,342 for staining the nuclei. A scale bar representing 50 μm was included. The nuclear translocation was quantified using Pearson’s correlation coefficient and analyzed using Image J software with the JACoP plugin. (c) The verification of β-catenin translocation was accomplished through the separation of cytoplasmic and nuclear fractions. Western blot analysis of β-catenin was conducted on these fractions, employing Lamin B1 and α-tubulin as internal controls for nuclear and cytoplasmic compartments. The presented data represents the mean ± S.D. from three independent experiments. Statistical significance was determined by unpaired t-tests or One-way ANOVA, followed by Dunnett’s post-hoc analysis. Significance levels were denoted by asterisks (* for P < 0.05, ** for P < 0.01). These analyses were carried out to compare the results with the DMSO control. The original images for western blot analysis were shown in “Supplementary information”.
