SAG suppressed the growth of FARAGE and GM12878 cells. (a) IC50 of SAG in the indicated cell lines. (b) CCK-8 assays showed the growth of FARAGE and GM12878 cells upon treatment with SAG at concentrations of 5 μM, 10 μM, 20 μM, and 40 μM for 48 h. The OD450 value was measured. (c) BrdU assays showed the growth capacity of FARAGE and GM12878 cells following treatment with SAG at concentrations of 5 μM, 10 μM, and 20 μM for 48 h. The red panel indicates BrdU. (d) Quantification of panel C, the percentage of BrdU positive cells are shown. Scale bar, 100 μm. The percentage of BrdU positive cells was measured. ***p < 0.001. CCK-8, cell counting kit-8; PBMC, peripheral blood mononuclear cell; SAG, Sanguinarine.
Fig 2.
SAG stimulated cell cycle arrest in FARAGE and GM12878 cells. Flow cytometry assays demonstrated the cell cycle difference in FARAGE (up) and GM12878 (down) cells upon treatment with SAG at concentrations of 5 μM, 10 μM, and 20 μM for 48 h. The percentage of cells at G1, G2/M, and S phases was calculated and compared. ***p < 0.001. SAG, Sanguinarine.
Fig 3.
SAG stimulated cell apoptosis in FARAGE and GM12878 cells. (a) Flow cytometry assays showed the apoptosis degree of FARAGE (up) and GM12878 (down) cells upon treatment with SAG at concentrations of 5 μM, 10 μM, and 20 μM for 48 h. The percentage of apoptosis cells was calculated and compared. (b) Immunoblot analysis showed the expression of Bax and cleaved caspase-3 in FARAGE (left) and GM12878 (right) cells after treatment with SAG at concentrations of 5 μM, 10 μM, and 20 μM for 48 h. The relative expression of Bax and cleaved caspase-3 is shown. *p < 0.05, **p < 0.01, ***p < 0.001. SAG, Sanguinarine.
Fig 4.
SAG restrained the Wnt/beta–catenin axis in FARAGE and GM12878 cells. Immunoblot analysis showed the expression of Axin, c-Myc, and beta-catenin in FARAGE (left) and GM12878 (right) cells upon treatment with SAG at concentrations of 5 μM, 10 μM, and 20 μM for 48 h. The relative expression levels of Axin, c-Myc, and beta-catenin are shown. *p < 0.05, ***p < 0.001. SAG, Sanguinarine.