Fig. 5.

IDOL-mediated LDLR degradation is dynamin independent. A, B: HepG2 cells were cultured for 16 h in sterol-depletion medium to stimulate LDLR expression. Subsequently, cells were treated with 1 μM GW3965 or vehicle in the presence (+) or absence (−) of 80 μM dynasore for 4 h. A: Cells were incubated for 30 min with 5 μg/ml DyLight 488-labeled LDL. LDL uptake in vehicle-treated cells was set to 100% (n = 4, *P < 0.01 from vehicle treated cells). B: Total cell lysates were immunoblotted as indicated. C: HeLa cells were transfected with 20 nM control or DNM1/2 siRNA and subsequently incubated for 16 h with sterol-depletion medium to induce LDLR expression. Cells were then treated for 6 h with vehicle or 1 μM GW3965. Immunoblots are representative of three independent experiments.