Figure 4.

Characterizion of the effects of Rm-HE on apoptotic signaling mediators. (A) Effect of Rm-HE on the expression and cleavage of apoptosis-related proteins in Jurkat cells. Expression of the anti-apoptotic proteins Bcl-XL, Mcl- 1 and Bcl-2 was analyzed in extracts of Jurkat cells treated as indicated by immunoblot with specific antibodies. Tubulin was used as an internal control. (B) Expression of pro-apoptotic proteins Bim, Bax and Bad was analyzed in extracts of Jurkat cells treated as indicated by immunoblot with specific antibodies. Tubulin was used as an internal control. (C) Fas Ligand induction was analyzed in extracts of Jurkat cells treated as indicated by immunoblot with specific antibodies. Tubulin was used as an internal control. (D) Statistical analysis for Fas-L expression blots in Rm-HE treated cells. Results indicate the average fold increase ± S.E.M in Fas-L expression relative to untreated cells from three independent experiments. The difference between untreated cells and cells treated for 16 h with Rm-HE are statistically significant (Student’s t-test: **P < 0.01). (E) Effect of Rm-HE on Jurkat cell viability in the presence of a JNK inhibitor. Jurkat cells were pre-incubated for 1 h with 10μMSP600125 and then 40 μg/ml of Rm-HE were added for 24 h and 48 h, as indicated. Cell viability is represented as a percentage relative to untreated cells. Data is means ± S.E.M. from three independent determinations performed in duplicate. (F) Effect of Rm-HE on the JNK phosphorylation in Jurkat cells. Expression of JNK and phospho-JNK were analyzed in extracts of Jurkat cells treated as indicated by immunoblot with specific antibodies. Tubulin was used as an internal control.