Figure 5. ERAD activity is enhanced by R1881.

LNCaP cells were transfected with (A) CD3δ and (B) KAI1 on 100 mm dishes. Six hours later, cells were splitted onto 6 well dishes. Next day, cells were first androgen-starved and then treated with R1881. Cycloheximide was added into indicated samples 18 h and 21 h after R1881 treatment and cells were harvested 24 hours after R1881 addition. The level of CD3δ and KAI1 was detected by immunoblotting against their tags HA and myc, respectively and quantified by normalizing samples to actin levels. The degradation rates of substrates (right) were analyzed using three independent experiments.