CG&D
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cell Growth & Differentiation

Right arrow Help viewing high resolution images
Right arrow Return to article
Click on image to view larger version.



Fig. 3. The effect of heme deficiency on p44/42 MAPK (ERK1/2) phosphorylation during NGF-induced PC12 cell differentiation. A, a time course of ERK1/2 activation by NGF. PC12 cells were treated with no reagent (N, Lane 1), or succinyl acetone alone (SA, Lane 2), or NGF alone (NGF, Lanes 3–7). Proteins were extracted from PC12 cells induced with NGF for 5 (Lane 3), 10 (Lane 4), 30 (Lane 5), 60 (Lane 6), or 180 min (Lane 7). The same amount of total cellular proteins was loaded in each lane. Activated ERK1/2 proteins were detected by Western blotting with an antibody specific for phospho(Thr-202/Tyr-204)-p44/42 (ERK1/2). The positions of phospho-p44/42 (p-p44/42) and p44/42 are marked. B, the total ERK1/2 protein level in NGF-induced cells. Total cellular ERK1/2 proteins were detected by using an antibody specific for both phosphorylated and nonphosphorylated p44/42 (ERK1/2). The same blot in A was stripped and reprobed here. C, a time course of ERK1/2 activation by NGF in heme-deficient cells. PC12 cells were treated with no reagent (N, Lane 1), or succinyl acetone alone (SA, Lane 2), or NGF and succinyl acetone (NGF+SA, Lanes 3–7). Proteins were extracted from PC12 cells induced with NGF for 5 (Lane 3), 10 (Lane 4), 30 (Lane 5), 60 (Lane 6), or 180 min (Lane 7). The same amount of total cellular proteins was loaded in each lane. Activated ERK1/2 proteins were detected by Western blotting with an antibody specific for phospho(Thr-202/Tyr-204)-p44/42 (ERK1/2). D, the total ERK1/2 protein levels in NGF-induced heme-deficient cells. The same blot in C was stripped and reprobed with an antibody specific for both phosphorylated and nonphosphorylated p44/42 (ERK1/2) here. E, the effect of heme add-back on ERK1/2 phosphorylation. PC12 cells were treated with no reagent (Lane 1), NGF alone (Lanes 2–4), NGF and heme and succinyl acetone (Lanes 5–7), or NGF and succinyl acetone (Lanes 8–10). Proteins were extracted from PC12 cells induced with NGF for 10 (Lanes 4, 7, and 10), 30 (Lanes 3, 6, and 9), or 60 min (Lanes 2, 5, and 8). Activated ERK1/2 proteins were detected by Western blotting with an antibody specific for phospho(Thr-202/Tyr-204)-p44/42 (ERK1/2). F, the total ERK1/2 protein levels in NGF-induced heme-sufficient and -deficient cells. The same blot in E was stripped and reprobed with an antibody specific for both phosphorylated and nonphosphorylated p44/42 (ERK1/2) here.





Right arrow Return to article


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cell Growth & Differentiation