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. 2021 Jun 1;12(15):4626–4637. doi: 10.7150/jca.59740

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HOXA5 mediates invasion and migration abilities and stemness of breast cancer cells and induces aggressive phenotypes. (A) Matrigel invasion assay in parent MCF7, TAMR, and TAMR cells transfected with control or HOXA5 siRNAs. Representative images acquired by fluorescent microscopy after DAPI staining are shown. Quantitative interpretation was attained using ImageJ. (B) Migration assay in parent MCF7, TAMR, and TAMR cells transfected with control or HOXA5 siRNAs. Representative images acquired by fluorescent microscopy after DAPI staining are shown. Quantitative interpretation was attained using ImageJ. (C) Western blotting for epithelial and mesenchymal markers in parent MCF7, TAMR, and TAMR cells transfected with control or HOXA5 siRNAs. (D) Left panel shows representative microscopic images of breast cancer stem cell (BCSC) sphere growth of parent MCF, TAMR, and TAMR cells transfected with control or HOXA5 siRNAs. Right panel shows graphic presentation of the average sphere diameter. The data were retrieved and analyzed on the 14^th day of spheroid culture. (E) Reverse transcription-quantitative PCR analysis of SOX2, OCT4, and NANOG in parent MCF7, TAMR, and TAMR cells transfected with control or HOXA5 siRNAs. All experiments were performed in triplicate. *** p < 0.001 compared with MCF7 and siCON respectively by Student's t-test or one-way ANOVA.