FIGURE 2:

BCH domain–mediated differentiation of PC12 cells occurs via the Ras/MAPK pathway. PC12 cells transfected with BCH (A) and BPGAP1 (B) were made quiescent before treatment with dimethyl sulfoxide (DMSO; control) or U0126 (5 mm) either with or without EGF (100 ng/ml) for 48 h before they were processed by indirect immunofluorescence for confocal microscopy. (C) PC12 cells were cotransfected with BCH and Mek2-K101A, made quiescent, and stimulated with EGF (100 ng/ml) for 48 h before they were processed by indirect immunofluorescence for confocal microscopy. Red arrowheads point to the long bipolar neurites. The merged panel shows inhibition of BCH-mediated PC12 differentiation by Mek2-K101A with the white arrowheads pointing to lack of neurites. DIC, differential interference contrast. Scale bars, 20 μm. (D–F) Quantitative representation of the data depicted in A–C, respectively, as mean of n = 3, p < 0.01; error bars represent SEM.