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. 2016 May 18;7(44):71151–71168. doi: 10.18632/oncotarget.9463

Figure 1. FASN expression correlates with malignant progression of MCF10CA cells.

Figure 1

(A) Western blot analysis of FASN protein in cells from various tumor stages. NeoT: neoplasmic, kcl2: atypical hyperplasia, AT1: hyperplasia, DCIS: ductal carcinoma in situ, CA: metastatic/invasive cancer. (B) Inhibition of FASN reduces cell viability. CA1d and CA1a cells were treated with cerulenin or C75 and cell viability was assessed by MTT reduction. Dose–response curves were plotted as percentages of the control cells' absorbance (= 100%). Results from one representative experiment are presented as mean ± SD; p ≤ 0.02 for cerulenin and p ≤ 0.05 for C75, respectively. (C) FASN inhibition impairs anchorage-independent growth. CA1d and CA1a cells were grown in soft agar in the presence of increasing concentrations of cerulenin for 21 d. Colony numbers from one representative image are shown as mean ± SD; *p < 0.05. (D) Representative images from a soft agar growth assay as described in C. (E) FASN knockdown efficiency in stably-transduced CA1d cells; WT = wild type. (F) Depletion of FASN decreases cell viability. Control or FASN-depleted cells were assessed by MTT reduction 72 h post transfection and the results from one representative experiment are presented as mean ± SD; *p ≤ 0.05. (G) FASN depletion inhibits anchorage-independent growth. FASN-depleted or control CA1d cells were grown in soft agar and colony numbers from one representative experiment are plotted as mean ± SD, *p < 0.01. (H) Inhibition of FASN increases the levels of energy stress markers. CA1d cells were treated with either cerulenin or C75 and cell lysates were subjected to western blotting. Note a substantial increase in p-AMPK and p21 upon FASN inhibition. (I) Depletion of FASN increases the levels of energy stress markers. Control and FASN-depleted CA1d cells were lysed and subjected to western blotting. (J) Palmitate increases cell viability of FASN-depleted cells. CA1d/vector control or FASN-depleted cells were exposed to increasing concentrations of palmitate-BSA in 0.5% HS containing medium and cell viability was assessed by MTT reduction 72 h after treatment. Data are presented as mean ± SD from two independent experiments.