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. 2004 Nov 8;167(3):479–492. doi: 10.1083/jcb.200403093

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Copyright © 2004, The Rockefeller University Press

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Figure 5. — Effects of FAIM on ERK pathway stimulation. (A) PC12 cells stably transfected with empty vector or with FAIM-S were treated with NGF (100 ng/ml) or with NGF plus 50 μM of PD98059 for 1 d. Histogram shows the neurite length measurements of the cultures transfected with the indicated cDNA. Significant differences are indicated (*, P < 0.05; **, P < 0.001; t test). (B) PC12 cells stably transfected with empty vector (N) or FAIM-S (S) were serum starved for 12 h, pretreated (+) or not (−) with 50 μM PD98059 for 30 min, and stimulated (+) or not (−) with 100 ng/ml NGF for 5 min. Protein extracts were analyzed by Western blotting with an anti–phospho-ERK antibody (top) and stripped and reprobed with anti–pan-ERK and anti-tubulin antibodies (middle) to control the protein loading. Anti-FLAG immunoblotting (bottom) confirmed FAIM-S overexpression in transfected PC12 cells. White lines indicate that intervening lanes have been spliced. (C) PC12 cells stably transfected with empty vector (N), FAIM-S (S), or RNAi2 (R) were serum starved for 12 h and stimulated with 100 ng/ml NGF for the indicated times. Protein extracts were processed as in B.