BXL0124 inhibits STAT3 activation and CD44-STAT3 interaction in MCF10DCIS cells.

<p>(A) MCF10DCIS cells were treated with BXL0124 (10 nM) for 24 h. Cells were fixed using 4% paraformaldehyde and stained with antibody against pSTAT3 (green). Nuclei were stained with To-PRO-3 (blue). (B) MCF10DCIS cells were treated with BXL0124 (0.1, 1 or 10 nM) for 24 h. Each cell lysate was incubated with oligonucleotides containing STAT3 binding sequences. The amount of STAT3 bound to the oligonucleotides was measured as chemiluminescent intensity value by luminometer. The fold change of chemiluminescent intensity value in each sample from control was determined (*p<0.05). (C) and (D) MCF10DCIS cells were treated with BXL0124 (10 nM) for 24 h, then the cell lysates were immunoprecipitated with STAT3 or JAK2 antibodies. The amounts of given proteins interacting with STAT3 or JAK2 were determined by Western blot analysis. STAT3 and JAK2 were used as loading control for each immunoprecipitation experiment, respectively.