Figure 2. Ternatins inhibit global protein synthesis.
(a) HCT116 cells were treated with compound 4 (five-fold dilutions) for 5 hr before labeling with 35S-Met for 1 hr. Cell lysates were separated by gel electrophoresis. Newly synthesized and total proteins were visualized by autoradiography and Coomassie staining. (b) Cells were treated as in (a), and 35S-labeled proteins were quantified by liquid scintillation counting after TCA precipitation (mean ± SEM, n = 3). (c) Left panel: HeLa cells were treated with 4 (5 μM) or DMSO for 20 min. Right panel: HeLa cells were treated with 4 (5 μM), cycloheximide (CHX, 100 μg/mL), or DMSO for 15 min, followed by harringtonine (HT, 2 μg/mL) for 20 min. After compound treatment, lysates were fractionated on 10–50% sucrose density gradients with absorbance detection at 254 nm.