Effects of crude extracts of Pseudocyphellaria granulata and physciosporin on cancer stemness-related protein and gene expression. (A) Western blot analysis of cancer stem cell markers aldehyde dehydrogenase-1 (ALDH1), cluster of differentiation 133 (CD133), CD44, Lgr5, and Musashi-1 in CSC221 cells treated with acetone extracts of P. granulata or various concentrations of physciosporin. (B,C) Quantitative analysis of the protein expression levels of cancer stem cell markers ALDH1, CD133, CD44, Lgr5, and Msi-1 in CSC221 cells treated with various concentrations of acetone extracts of P. granulata (B) or physciosporin (C). (D,E) Relative mRNA expression of cancer stemness markers ALDH1, CD133, CD44, Lgr5, Msi-1, Bmi-1, EphB-1, Hes-1, Gli-1, Gli-2, and SMO on CSC221 cells after treatment with various concentrations of acetone extract of P. granulata (D) and physciosporin (E). Quantitative data were obtained from at least two independent experiments. Data are means ± standard error of the mean (SEM), and statistical analysis was performed by one-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001 vs. DMSO-treated CSC221 cells.