Figure 1.

Measurement of total 4EBP1 in AML cell lines. A) Electropherogram depicting levels of total and phosphorylated 4EBP1 in AML cell lines. AML cell lines MV4-11, MOLM-14, OCI-AML3 and HL60 were analyzed at baseline for expression of 4EBP1. 80 ng of protein was used for analysis. β-2 Microglobulin was used as loading control. Total 4EBP1 antibody detects both phosphorylated and non-phosphorylated protein. Samples were treated with lambda-phosphatase or reaction buffer alone and decrease in phosphorylation was noted. X-axis represents iso-electric pH and y-axis represents luminescence units. B) Change in phosphorylation was measured at exposure times varying between 30–960 secs after phosphatase treatment in all cell lines. The experiments were performed in triplicate (*p < 0.05).