Fig. 5

Treatment with IC₅₀ of salinomycin and the analogs 2a-c result in similar effects on CSCs and cell proliferation. a The CD44⁺/CD24⁻ population evaluated by flow cytometry after 72 h of treatment. b The ALDH⁺ population evaluated by flow cytometry after 72 h of treatment. c Colony forming efficiency evaluated using a serum free soft agar assay. The cells were treated for 72 h and then reseeded at cloning density. The colonies were counted after 2 weeks of incubation. d Colonies obtained from the serum free soft agar assay. Bars = 40 μm. e Cell cycle phase distribution evaluated by flow cytometry. f Representative Western blots used for densitometric scanning. g–h Expression of cyclin A2 and p27, respectively. Data are represented as mean ± SEM for n ≥ 3. The columns in (a–d) show mean ± SEM for n = 4. *P < 0.05, **P < 0.01, ***P < 0.001. ns: no significant difference. SA: salinomycin, 2a: carbamate, 2b: acetate, 2c: carbonate and 3: C1-methyl ester