Fig. 5
From: BRD4/nuclear PD-L1/RelB circuit is involved in the stemness of breast cancer cells
RelB/p65 complex regulated mammosphere formation. A CoIP assay. CoIP assay was performed in mammospheres derived from MDA-MB-231 cells. The cell lysate was incubated with the RelB antibody or corresponding IgG. The immunoprecipitants were blotted with p65 or p50. The RelB/p65 complex was verified through the IP assay of p65. B Mammosphere formation regulation of p65. Cultured MDA-MB-231 cells were transfected with si-p65 for 2 days. Mammospheres derived from si-p65-transfected cells were incubated for 7 days. p65 knockdown was verified by Western blot, and mammosphere images (right) were taken at ×10 magnification. C Inhibitory effect of p65 inhibitor caffeic acid phenethyl ester (CAPE) in triple-negative breast cancer cells and mammospheres. The proliferation of CAPE-treated cells was measured using the MTS assay. The cells were treated with increasing concentration range of CAPE for 24 h. Mammospheres derived from MDA-MB-231 cells were given indicated concentrations of CAPE for 7 days. The RelB-inhibitory effect of CAPE was verified by Western blot of nuclear fraction. 20 µM CAPE was used to perform Western blot. Experiment values are represented as the mean ± SD of triplicates. Compared with the control as determined by student’s t-test or one-way ANOVA with Dunnett’s multiple comparisons tests, * p < 0.05