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Fig. 5. The effect of heme deficiency on CREB activation during NGF-induced PC12 cell differentiation. PC12 cells were treated with no reagent (N, Lanes 1 and 8), or succinyl acetone alone (SA, Lanes 2 and 9), or NGF alone (NGF, Lanes 3–7), or NGF and succinyl acetone (NGF+SA, Lanes 10–14). Proteins were extracted from PC12 cells induced with NGF for 5 (Lanes 3 and 10), 10 (Lanes 4 and 11), 30 (Lanes 5 and 12), 60 (Lanes 6 and 13), and 180 min (Lanes 7 and 14). The same amount of total cellular proteins was loaded in each lane. Protein extracts were subjected to Western blotting analysis with antibodies against phospho(Ser-133)-CREB (A) or total cellular CREB (B).