FIGURE 3.

Suppression of P-Bad(Ser-155, Ser-112) by M analogs. A, P-Bad(Ser-155) of M-treated Jurkat and COS-1 cells, determined by SDS-PAGE/Western blot analysis, in the presence or absence of 10 μm Forskolin (Fors) and 30 μm IBMX added 1 h prior to sampling, as indicated. The P-Bad(Ser-155)/Bad ratio of extracts of non-treated controls is defined as 1.0. Values are mean ± S.E. of three independent experiments. *, significant as compared with non-treated controls (p < 0.05). #, significant as compared with Forskolin/IBMX-treated controls (p < 0.05). Inset: representative blots. B, cAMP content of Jurkat and COS-1 cells treated as described in A. The cAMP content of extracts of non-treated controls is defined as 1.0. Values are mean ± S.E. of four independent experiments. *, significant as compared with non-treated controls (p < 0.05). #, significant as compared with Forskolin/IBMX-treated controls (p < 0.05). C, percent TMRM fluorescence intensity of M-treated Jurkat cells in the presence or absence of 500 μm Bt₂cAMP added 4 h prior to sampling, as indicated. The mean TMRM fluorescence intensity of non-treated controls is defined as 100%. Values are mean ±S.E. of five independent experiments. *, significant as compared with non-treated controls (p < 0.05). #, significant as compared with Mαα-treated controls (p < 0.05). Inset: representative histogram of non-treated controls (a) and cells treated with Bt₂cAMP (b), M (c), or M + Bt₂cAMP (d). D, cAMP content of M-treated Jurkat cells in the presence and absence of 5 μg/ml cholera toxin (CTX) (left ordinate, mean of two independent experiments differing by no more than 10%) or 50 ng/ml pertussis toxin (PTX) (right ordinate, mean ± S.E. of three independent experiments) added 3 h prior to sampling, as indicated. The cAMP content of extracts of non-treated controls is defined as 1.0. *, significant as compared with non-treated controls (p < 0.05); #, significant as compared with PTX-treated controls (p < 0.05). E, Bad, P-Bad(Ser-112), ERK, and P-ERK(Y204) of M-treated Jurkat cells, determined by SDS-PAGE/Western blot analysis, in the presence or absence of 50 nm PMA added 1 h prior to sampling. The -Bad(Ser-112)/Bad and P-ERK(Y204)/ERK ratio of extracts of non-treated controls is defined as 1.0. Values are mean ± S.E. of three independent experiments. *, significant as compared with non-treated controls (p < 0.05); #, significant as compared with PMA-treated controls (p < 0.05). Inset: representative blots.