Figure 2

There was a negative correlation between JWA and topoisomerase IIα protein expression.

(a) and (b) Expression of topoisomerase IIα was elevated and that of topoisomerase IIα was reduced in lung cancer tissues compared with those in matched normal tissues. β-Tubulin expression was served as a loading control. JWA and topoisomerase IIα expressions in cancer tissues (T) and paired non-cancerous normal tissues (N) of lung cancer patients were analyzed by Western blotting. The level of each protein was normalized against β-tubulin. NCI-H460 cells were transiently transfected with Flag-JWA or Flag- topoisomerase IIα plasmid (1–4 μg). Whole-cell extracts were prepared 24 h after transfection and the expression of target proteins and mRNAs was examined. As analyzed by western blot assay, topoisomerase IIα levels were decreased after transfection with JWA plasmid (c) Also, overexpression of topoisomerase IIα does dependently inhibited JWA protein expression (d) β-actin was used for the protein loading control. Total RNAs were isolated from NCI-H460 cells transfected with Flag-JWA or Flag- topoisomerase IIα plasmid (4 μg) and subjected to real-time PCR. JWA transfection suppressed topoisomerase IIα mRNA expression (e) The level of JWA mRNA was down-regulated in topoisomerase IIα overexpressed NCI-H460 cells (f) GAPDH was used as an internal control. Error bars represent the mean ± SD of triplicate experiments. Statistical differences to the controls were shown as *p < 0.05, **p < 0.01.