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. 2011 Aug 22;6(8):e23850. doi: 10.1371/journal.pone.0023850

Figure 5. Aconitase inhibition interferes with EPO activation of ERK signaling in a pathway-specific manner.

Figure 5

(A) Steady state phosphorylation of ERK and RSK as determined by immunoblotting of whole cell lysates of cells from days 3–4 erythroid cultures treated with 50 µM FA and 10 µM U0126 as indicated (NT, untreated). (B) Time course of ERK1/2 and RSK phosphorylation. Cells from day 3 erythroid cultures ±50 µM FA were subjected to 3 hours of cytokine starvation followed by stimulation with 4.5 U/ml EPO for the indicated time periods. (C) EPO induction of multiple downstream pathways in cells treated as in panel B. (D) Densitometric analysis of pERK1/2:ERK1/2 and pRSK:RSK levels in five independent experiments conducted as in panel C. Presented are mean values ± SEM for relative phosphoprotein signal divided by total protein signal; * P<0.05; † P<0.01; n.s., not significant.