Figure 5.

1-Heptanol inhibits phosphorylation of SAPK/JNK (JNK) but not ERK and p38 in antigen-activated BMMCs. (A–F) IgE-sensitized BMMCs were pre-incubated with vehicle (BSS-BSA; Control) or 2.5 mM 1-heptanol for 15 min and then activated or not with antigen (TNP-BSA; 0.5 µg/mL) for the indicated time intervals. Whole-cell lysates were size fractionated by SDS-PAGE and analyzed by immunoblotting with phosphotyrosine-specific antibodies recognizing ERK^Y204 ((A,B); n = 3), p38^T180/Y182 ((C,D); n = 3), and JNK^T183/Y185 ((E,F); n = 3). Antibodies specific for the corresponding proteins were used as loading controls. Representative immunoblots with the corresponding protein loading controls are shown in (A,C,E). The results of quantification of the tyrosine-phosphorylated proteins in activated cells are normalized to the signals in non-activated cells and loading control proteins (B,D,F). Values are mean ± SEM calculated from the numbers indicated above (n), which indicate the numbers of biological replicates. Statistical significance of intergroup differences is indicated in (F); **** p < 0.0001. Numbers on the left in (A,C,E) indicate positions of molecular weight markers in kDa.