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. 2013 Oct 14;8(10):e77060. doi: 10.1371/journal.pone.0077060

Figure 2. Effect of ACSL4 expression on breast cancer growth .

Figure 2

A doxycycline-inducible line of MCF-7 cells was transfected with either a control or ACSL4-expressing plasmid as described in the text. Both control and ACSL4-MCF-7 cells were treated with 1μg/ml of doxycycline. (A) RT-PCR analysis of mRNA for ACSL4, AR, ER and PgR in vector control and ACSL4-MCF-7 cells. GAPDH was used as a loading control. (B) Immunoblot analysis of whole cell lysates showing expression of ACSL4 and ER with β-actin as loading control. The increase in ACSL4 is 163 fold, and the decrease in ER is 68%. (C) Comparison of proliferation of control with ACSL4-MCF-7 cells grown in phenol red-free medium supplemented with charcoal-stripped FBS. Values shown are the means of triplicate determinations ± 1SD. The difference between the curves is significant, p<0.0001. (D) The effect of estradiol on proliferation of doxycycline-induced ACSL4-transfected MCF-7 cells grown as in (C). Values shown are the means of triplicate determinations ± 1SD. The difference between the curves is significant, p<0.0001. (E) The effect of estradiol on proliferation of doxycycline-treated control MCF-7 cells grown as described above for panel C. Values shown are the means of triplicate determinations ± 1SD. The difference between the curves is significant, p<0.0001.