Figure 1: PDK1 regulates protein synthesis in CD34⁺ derived, cultured, human megakaryocytes.

A. CD34+ megakaryocytes were left alone or treated with BX-795 (1μM) from culture day 11 – 13. Cells were collected on Day 13 and adhered to fibrinogen in media that contains S³⁵ methionine for 2 hours. Cyclohexamide (Cyclo) was used as a positive control. Protein synthesis was measured using scintillation counts. B. On culture day 13, CD34+ megakaryocytes were adhered to a fibrinogen coated surface for 60 min in the presence or absence of BX-795 (1μM). Cell lysates were immunoprecipitated with 4EBP1 and samples were probed for eIF4E using an anti-eIF4E mouse mAb. C. CD34+ megakaryocytes were left alone or adhered to fibrinogen (60 min) in the presence or absence of BX-795. Cells were lysed and incubated with m7 GTP- agarose beads. The beads were eluted to collect the eIF4E protein. Samples were analyzed for eIF4E binding using western blot (**P < 0.05) (***P < 0.01).