Figure 4

Characterizing extracellular PPIA-mediated activation of ERK1/2 phosphorylation within multiple cell lines. A) ERK1/2 phosphorylation in PPIA stimulated HEK293T, MOLM13, PANC-1, and L3.6pL cells was determined by immunoblotting. Cells were treated with either buffer alone (−) or 25 μM PPIA (+) for 10 min and stored in SDS loading buffer for subsequent immunoblotting analysis of pERK1/2 and total ERK1/2 as a control. B) PPIA-induced time-dependent induction of ERK1/2 phosphorylation as monitored by flow cytometry is shown for both MOLM13 and PANC-1 cells for the indicated time points also stimulated with 25 μM PPIA. The data shown represents the mean of fluorescence intensity (MFI) for each time point measured in triplicate and normalized to the average initial MFI.