HeLa/TSA was up-expressed UbB and exhibited higher ability of the mammosphere formation.

<p><b>A</b>, HeLa/TSA cells were analyzed for UbB, UbC, UbA52 and UbA80 mRNA levels. The columns represent the average of three separate experiments; error bars, SD; *, <i>p</i><0.05. <b>B</b>, HeLa/TSA cells were analyzed for the protein levels of Ub proteins, pSTAT3, Vimentin, pAKT and Mdr-1, β-actin was used as a loading control. <b>C</b>, Quantitative analysis of the proliferation index determined by the CCK-8 assay for the indicated times. The results shown are averaged through three separate experiments; error bars, SD. <b>D</b>, Representative examples of tranwell assay were photographed at 200× magnifications. <b>E</b>, Themorphologic appearance of a representative mammosphere for the indicated times was photograghed at 200×magnifications. <b>F</b>, Spheroid formation assays were performed by limiting dilution with 256 cells to one cell per well of a 96-well Ultra-Low Adherent plate. This experiment was performed for triple times with six wells per well dilution. The number of colonies was counted at day 14. <b>G</b>, Mammosphere cells of HeLa and HeLa/TSA were re-cultured in an ordinary adherent plate for 24 h, and then treated with TSA, DDP, and PTX for 48 hours or UV for 5 min. The apoptosis cells were quantified using Annexin V/PI staining and the flow cytometry analysis. The values are average and SD (error bars) of 3 separate experiments, *, <i>p</i><0.05. <b>H</b>, HeLa/TSA mammosphere cells were analyzed for UbB, UbC, UbA52 and UbA80 mRNA levels. GAPDH was used as a reference control.