Induction of Tumor Cell Death through Targeting Tubulin and Evoking Dysregulation of Cell Cycle Regulatory Proteins by Multifunctional Cinnamaldehydes
Figure 7
Interaction of cinnamaldehydes with sulfhydryl groups in tubulin and their effects on tubulin organization in HCT 116 cells.
(a) Cinnamaldehydes were incubated with purified tubulin for 30 min at 37°C and aliquots of samples were labeled with BIAM. Western blotting by HRP-streptavidin was performed to detect BIAM labeling of free thiol groups. Equal amount of tubulin across all samples was ascertained by probing for β-tubulin. (b) Representative immunofluorescence microscopic images of cells treated with DMSO (i & ii), 30 µM BCA (iii & iv), 30 µM FHCA (v & vi), and 30 µM FBCA (vii & viii). Cells cultured on coverslips and treated with DMSO or cinnamaldehydes for 8 h were fixed, and subjected to immunocytochemical analysis using anti-β-tubulin antibody. Coverslips were then mounted on slides using mounting medium containing DAPI and images were obtained at 100×magnification.