Combination effect of salinomycin and STAT3 inhibitors on apoptosis, migration, and CD44⁺/CD24^- stem-like population.

<p>(A) Cells were treated with salinomycin (2 μM, 48 h) and/or S3I-201 (50 μM) or LLL12 (1 μM) and annexin V/PI analysis was performed. Quantitative graph of annexin V-positive population is shown (bottom panel, *** <i>p</i><0.001 and ### <i>p</i><0.001). (B) Effect of combined treatment with salinomycin and S3I-201 on protein expression of STAT3, phospho-STAT3, cyclin D1, and apoptosis-related factors. (C) Cells were co-treated with salinomycin (2 μM) and S3I-201 (50 μM) or LLL12 (1 μM) and the relative wound density was assessed over 24 h. Quantitative graph of relative migration is presented (** <i>p</i><0.01 and ## <i>p</i><0.01). (D) Effect of salinomycin (2 μM, 48 h) combined with S3I-201 (50 μM) or LLL12 (1 μM) on the CD44⁺/CD24^- stem-like population. The graph represents the percentage of CD44⁺/CD24^- cells (bottom panel, * <i>p</i><0.05 and ## <i>p</i><0.01). The results were expressed as mean ± SEM and analyzed by one-way ANOVA followed by Bonferroni’s <i>post hoc</i> test (* <i>p</i><0.05, ** <i>p</i><0.01, and *** <i>p</i><0.001, versus DMSO control; ## <i>p</i><0.01 and ### <i>p</i><0.001, versus salinomycin only). All experiments were independently performed at least three times (n = 3).