Fig. 2

SAH5-EJ1 enters cells, colocalizes with mitochondria, and survives in vivo, resulting in glioblastoma-derived cell death and glioma mouse model survival. A–C BT20 cells treated with Cy5.5-dye. A’ Cy5.5-only treatment, no SAH5-EJ1. B’ 10 µM Cy5.5-SAH5-EJ1. C’ 20 µM Cy5.5-SAH5-EJ1. Cells were incubated with Cy5.5 ± conjugation to SAH5-EJ1 (red) and mounted in DAPI (blue). Scale bars represent 50 µm. D–F Colocalization of SAH5-EJ1 with mitochondria in BT20 breast cancer cells. Cells were incubated with Cy5.5-SAH5-EJ1 (green) and MitoTracker Red (red). Single prime (‘) images represent single channel Cy5.5-SAH5-EJ1, double prime (“) images represent single channel MitoTracker Red. Arrowheads highlight changes in mitochondrial appearance. Scale bars represent 10 µm. G Mice were treated with Cy5.5-dye ± conjugation to SAH5-EJ1 and imaged after 30 min. Left mouse was treated with Cy5.5 dye through intravenous (IV) tail-vein injection. Middle mouse received Cy5.5-conjugated SAH5-EJ1 through subcutaneous (SC) injection in the right flank. Right mouse was treated with Cy5.5-conjugated SAH5-EJ1 through tail-vein (IV) injection. Experiment done in duplicate and representative images selected. Radiance scale provided on the left. H Luminescence distribution throughout organs. Cy5.5 dye only (left) versus Cy5.5-conjugated SAH5-EJ1 (right). B brain, H heart, Lu lung, K kidney, Li liver, S stomach. Radiance scale provided on left. I Cell viability assay performed in two glioblastoma-derived human cell lines (solid versus dashed) over 2 days comparing treatment with control peptide (CP) (grey lines) to SAH5-EJ1 (red lines). J Kaplan–Meier curve of glioblastoma mouse model treated with intravenous control peptide (blue) or 10 µg/g SAH5-EJ1 (red). Data shown represents mean. N = 6 for control, N = 4 for SAH5-EJ1. At the time of the GBM study, tumors formed when compound was limiting. Therefore, while enough compound was present for 4 mice, any extra mice were placed into the control arm