Figure 4.

Hormonal control of n-myc downstream regulated protein 1 (NDRG1)-Thr^346/356/366 phosphorylation in H441 cells. Experiments were undertaken using a strictly paired experimental design in which cells on 6 well plates were cultured for 5–6 days in either nominally hormone-free medium (hormone-deprived cells) or in medium supplemented with 0.2 µM dexamethasone (Dex.-treated cells). Before the cells were lysed to allow cellular protein to be extracted, the hormone-deprived cells were acutely stimulated with 0.2 µM dexamethasone (2 h) or 20 nM insulin (30 min), while the dexamethasone-treated cells were either untreated or acutely exposed to 20 nM insulin (30 min). (A) Typical Western blot illustrating the effects of these manoeuvres upon the cellular abundance of total and Thr^346/356/366-phosphorylated NDRG1. (B) Pooled data (n= 5) showing changes in the abundance of Thr^346/356/366-phosphorylated and total NDRG1 evoked by these experimental manoeuvres. *P < 0.05, value greater than that measured in hormone-deprived cells; ^†P < 0.05, value lower than that measured in hormone-deprived cells.