Fig. 6.

Long-chain unsaturated but not saturated FAs relieve the inhibition of TG synthesis imposed by Ubxd8. (A) HEK-293 cells were set up at 4 × 10⁵ cells/60 mm dish on day 0. On day 2 they were switched to medium supplemented with 10% delipidated FCS. After incubation for 16 h on day 3, the cells were labeled with 10 μM [¹⁴C]oleate in the absence or presence of 100 μM nonradiolabeled oleate for 4 h. The amount of [¹⁴C]oleate incorporated into TGs and DAGs was determined. (B) HEK-293 cells were set up at 1.5 × 10⁵ cells/60 mm dish on day 0. They were transfected with the indicated siRNA and incubated as described in Fig. 5A for the first three days. On day 4, these cells were labeled with 10 μM [¹⁴C]glycerol in the presence of indicated concentrations of nonradiolabeled oleate for 4 h. The amount of [¹⁴C]glycerol incorporated into TGs was determined. *, p < 0.05. (C) HEK-293 cells were set up and incubated as described in A for the first 2 days. On day 3 the cells were labeled with 10 μM [¹⁴C]oleate in the presence of 100 μM nonradiolabeled oleate (oleate) or 10 μM [¹⁴C]palmitate in the presence of 100 μM nonradiolabeled palmitate (palmitate) for 4 h. The amount of radio-labeled FAs incorporated into TGs and DAGs was determined. (D) HEK-293 cells were set up, transfected with indicated siRNA, and incubated as described in B for the first 3 days. On day 4, these cells were labeled with 10 μM [¹⁴C]palmitate in the presence of 100 μM nonradiolabeled palmitate for 4 h. The amount of [¹⁴C]palmitate incorporated into TGs and DAGs was determined. (A–D) Data are reported as mean ± S.E. from three independent experiments.