Figure 3.

Cyclin E levels decrease after 1 h of release from the DFO block. Confluent neuroblastoma cells were serum starved for 24 h then split and plated into media, one set with DFO (lanes 3–6, 9–12) and one with no DFO (lanes 1–2, 7–8) and were incubated for 24 h. After 24 h, one set of cells that was in DFO was replaced with CM (release). Medium in the others were replaced with CM (continuously in medium without DFO). The medium in all plates was aspirated 1 h later and the cells were lysed using 200 μL of hot SDS loading buffer. Forty microliter samples of the lysate were separated by 10% SDS‐PAGE and the proteins were transferred to PVDF membranes. This was first probed for cyclin E. Then the membrane was stripped and was probed for β‐actin. All treatments were conducted in duplicate.