Figure 6
A: Graph depicting the percentage of TrkA- or p75NTR- immunopositive cells in wild type (wt)PC12 cells and PC12 cell mutants lacking the low affinity NGF receptor (PC12p75NTR(-)) from flow cytometry data. Results shown are representative of 3 replicate flow cytometry experiments on the same cell line. B: SEAP release in the culture medium of PC12p75NTR (-) cells transfected for 24 hr with an NF-κB-sensitive SEAP reporter gene construct and treated for 12 hr with vehicle (Cont), 10 ng/ml NGF, 10 ng/ml TNFα or NGF plus TNFα (Both). Data are shown as mean ± S.E.M. from 3 independent replicate experiments. * : p < 0.05 vs. control or NGF-alone cells (two-tailed unpaired Student's t-test). C: Western blot detecting the presence of iNOS in wtPC12 cells and PC12p75NTR (-) cells treated for 24 hr with vehicle (Cont), 10 ng/ml NGF, 10 ng/ml TNFα or NGF plus TNFα (Both). Membrane was re-probed for β-actin (lower panel) to control for equal protein loading. Positive control (Pos) is 4 μg of total protein extracts from mouse macrophages. Results shown are representative of 4 replicate experiments. D: Western blot detecting the presence of TNFR-I in total protein extracts from wtPC12 cells and PC12p75NTR (-) cells. Twenty μg of total protein extracts from rat dorsal root ganglia (DRG) were used as a positive control.