Fig. 3

igure 3. DGAT1 antagonism decreases melanoma cell proliferation and survival (A) Confluence of cell lines transfected with DGAT1 targeting (007, 008, pool) or scrambled (sc) siRNAs (mean, n = 3, top). Corresponding protein expression of DGAT1 (bottom). (B) Left: relative cell number (mean, n > 3) in indicated cell lines determined by crystal violet following 72-h DGAT1 inhibitor treatment (50 μM AZD3988, 30 μM A922500, 50 μM AZD7687, or 70 μM T863). Right: percentage of cells in S-phase by using EdU incorporation following 24-h DGAT1 inhibitor treatment (mean, n > 3). (C) Relative cell number determined by crystal violet staining following 72-h A922500 treatment with cells grown in varying concentrations of fetal calf serum (FCS; mean ± SD, n > 3). (D) Relative cell number determined by crystal violet staining following 48-h A922500 treatment under normoxic or hypoxic conditions (1% O2) with cells grown in varying concentrations of FCS (relative to DMSO control for each condition; mean ± SD, n > 3). (E) Cleaved caspase index in indicated cell lines following transfection with either a DGAT1-targeting siRNA (007, 008, pool) or a sc control (mean, n = 3). (F) Protein expression of cleaved caspase-3 following treatment with/without A922500 for 24–72 h. (A) and (D) For significance: ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.