Figure 2.

Sesamin interacts physically with MCL-1, a predicted target molecule in T cells. (A) The flow chart how target proteins of sesamin were discovered from Genecards databank and swisstargetprediction analysis. (B) The structure of MCL-1 active domains (above) and molecular docking between BH3 domain of MCL-1 and sesamin by AMdock and pyMOL software. BH3-only protein Bim is marked as orange color following as sequence. A binding residue of sesamin at Arg 222 is marked as blue color. (C) Resting Jurkat T cells (1 X 10⁶) were lysed with RIPA buffer for pull-down assay. Lysates were incubated with control bead (Con bead) or/and bead conjugated with sesamin (SS-bead) and precipitated by centrifugation. Con beads and SS-beads were mixed in 100:0, 50:50, 0:100 ratios, respectively, to assess the dose-dependent enrichment of MCL-1 and Bak. Mcl-1 and Bak were detected from precipitated lysate by bead centrifugation (pull-down) and MCL-1 and β-actin were detected from whole lysate. The expression of detected MCl-1 from precipitated lysate by bead centrifugation (pull-down) was normalized with the expression of β-actin from whole lysate. Bar graph was presented as fold increase. All results are expressed as mean ± SEM of three independent experiments. Statistical comparisons among groups were performed using one-way ANOVA with Tukey's post hoc test. A p-value less than 0.05 was considered statistically significant (*, P < 0.05).