Figure 5.

Treatment with cryptolepine disrupts mitochondrial dynamics by activating AMPK phosphorylation, reducing Drp1 phosphorylation and inhibiting mTOR signaling in melanoma cells. (a) Immunofluorescence staining of p-AMPKα1/2 and p-Drp1 was performed after the treatment of cells with cryptolepine, as detailed in Materials and Methods. Photomicrographs were obtained using Keyence Fluorescence Microscope BZ-X710 (Keyence Corporation of America). A representative photomicrograph from each group is shown. (b) After treatment of A375 and Hs294t cells with cryptolepine for 24 h, total cell lysates were prepared and subjected to western blot analysis to determine the level of proteins involved in mTOR signaling. Blots were developed using a chemiluminescence-specific ECL system. Equal loading of proteins was verified by stripping the membrane and reprobing with anti-β-actin or vinculin antibodies.