Figure 7.

SR-IκBα transfection attenuates the NGF-induced differentiation and abolishes the effects of FAIM overexpression on neurite outgrowth. (A) PC12 cells stably transfected with empty vector (Neo) or with SR-IκBα. Total cell lysates were analyzed by Western blot using an antibody against IκBα (top). Membranes were stripped and reprobed with an antibody to α-tubulin to normalize the loading content per lane (bottom). (B–D) PC12 cells stably transfected with the empty vector (Neo) or with the vector expressing SR-IκBα and were treated with NGF for 5 min, TNFα for 15 min, or left untreated. Quantification of immunocytochemical detection of nuclear p65 is shown in B. Representative pictures of low and high magnification (arrows point to selected cells shown in insets) are shown in C. Bar, 25 μm. Quantification of neurite outgrowth was performed 1 d after NGF treatment (D). Asterisks indicate a significant (P < 0.001; t test) reduction of neurite outgrowth in PC12 cells expressing the SR-IκBα. (E) Neo or FAIM-S stably transfected cells were additionally transfected transiently with eYFP alone or with eYFP plus SR-IκBα. After 24 h, they were treated with NGF (100 ng/ml) for an additional 24-h period and neurite length was quantified. Significant differences are indicated (*, P < 0.001; t test).