Fig. 2.

ZBTB28 acts as a tumor suppressor to inhibit tumorigenesis of breast cancer. A Ectopic ZBTB28 expression at transcription and post-transcription level was detected by RT-PCR and Western blot. GAPDH was used as negative control. B Ectopic ZBTB28 expression impacts breast cancer cell growth was analyzed by the CCK-8 kit. ZBTB28 overexpression can suppress proliferation of breast cancer cells which were assessed through colony formation assay without soft-agar (C) and with soft-agar (D). E Flow cytometry analysis of cell cycle of MCF7 and MB231 which was overexpression pcDNA 3.1 or pcDNA-ZBTB28 by PI staining. F Ectopically expressed ZBTB28-induced apoptosis in breast cancer cells were tested by AO/EB staining assay. G Epithelial markers (E-cadherin, occludin) and mesenchymal markers (N-cadherin, snail) were detected via RT-PCR and Western blot, β-actin expression as control. H, I Transwell assay and wound-healing assay were used to investigate cell motility and invasiveness. Stem cell biomarker expression and growth of floating spheroid colonies were determined by RT-PCR (J) and spheroid formation assay (K) (independent experiment = 3, and a representative data was shown; ‘*’means p < 0.05; ‘**’means p < 0.01; ‘***’means p < 0.001)