The increasing effect of colony formation and drug-resistance in HeLa/TSA.

<p><b>A</b>, HeLa cells were treated with 500 nM TSA for the indicated times and subjected to analysis the mRNA level of UbB, UBC, UbA52 and UbA80 by real-time PCR and the corresponding protein level by western blotting. <b>B</b>, HeLa cells were incubated with TSA for 24 hours at a dose ranging from 200 to 800 nM and subjected to analysis the mRNA level of UbB, UbC, UbA52 and UbA80 by real-time PCR and the corresponding protein level by western blotting. <b>C</b>, Upper panel: representative dishes of the colony forming assay at day 7, 10 and 14. Lower panel: numbers of colony formed in HeLa and HeLa/TSA at day 7, 10 and 14. The columns represent the average of three separate experiments; error bars, SD; *, <i>p</i><0.05. <b>D</b>, HeLa/TSA cells were treated with TSA (1 μM), DDP (30 μM), and PTX (75 nM) for 48 h or UV, the apoptosis cells were quantified by Annexin V/PI staining and the flow cytometry analysis. The representative examples of the flow cytometry results were shown. <b>E</b>, Cells were treated as described in <b>D</b>, the average percentages of apoptosis cells were reported in the graphs. Values, mean percentages; error bars, SD; *, <i>p</i> < 0.05 (n = 3 replications).