ERα and PR receptor expression in MCF-7 Tet-Off empty vector and MCF-7 Tet-Off/ACSL4 human breast cancer cells.

<p>MCF-7 Tet-Off/ACSL4 cells were incubated with or without docicycline. After 96 h of doxycycline treatment, total RNA and cellular proteins from MCF-7 Tet-Off empty vector and MCF-7 Tet-Off/ACSL4 cells were extracted and subjected to RT-PCR analysis for ERα and PR and L19 mRNA expression and SDS-PAGE and Western blot analysis respectively. Semiquantitive RT-PCR of ERα and PR (panel A and B respectively). were performed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040794#s4" target="_blank">Materials and Methods</a>. The integrated optical density of PCR products was normalized with the corresponding L19 mRNA bands. Data represent mean ± SD of three independent experiments. Western blot analysis of ERα and PR (panel C and D respectively) were performed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040794#s4" target="_blank">Materials and Methods</a>. Using anti ERα and PR - and anti-β-tubulin antibodies, specific protein bands were detected in immunoblots by enhanced chemiluminescence. The integrated optical density of protein levels was quantified and normalized with the corresponding -tubulin signal. Data represent the means ± SD of three independent experiments. The asterisk indicates significant differences analyzed by Student’s test comparing MCF-7 Tet-Off empty vector and MCF-7 Tet-Off/ACSL4 cells, (**) <i>P</i>≤0.05.