Novel epoxy-tiglianes stimulate skin keratinocyte wound healing responses and re-epithelialization via protein kinase C activation (Supplementary Data)
Description
Epoxy-tiglianes are a novel class of diterpene esters isolated from a native Australian rainforest plant, Fontainea picrosperma (Euphorbiaceae). The prototype epoxy-tigliane, EBC-46 (tigilanol tiglate), possesses potent anti-cancer properties and is currently under clinical evaluation as a treatment for cutaneous and subcutaneous tumors in humans and domesticated animals. EBC-46 also stimulates exceptional dermal wound healing in vivo upon tumor destruction, manifested as accelerated wound re-epithelialization and closure. However, little is known how epoxy-tiglianes induce such favorable epithelial wound healing outcomes. The data presented provides supplementary information to our manuscript, demonstrating the stimulatory effects of EBC-46 and analogue, EBC-211, on the proliferative and migratory responses of human keratinocytes. Specifically, the data shows 1) The effects of both epoxy-tiglianes on the stimulation of accelerated cell cycle progression (assessed by Draq5 staining and Flow Cytometry analysis), 2) The effects of EBC-46 and EBC-211 on enhanced scratch wound repopulation (automated Time-Lapse Confocal Microscopy movies), in addition to epoxy-tigliane scratch wound movies in the absence and presence of anti-proliferative agent, mitomycin C (MC) and pan-protein kinase C (PKC) inhibitor, bisindolylmaleimide-1 (BIM-1). 3) We further provide Tables reporting the findings of Microarray analyses on the keratin (KRT); DNA synthesis/replication; cell cycle, proliferation and apoptosis; adhesion/migration; differentiation; proteinase; and cytokine/chemokine genes differentially expressed by EBC-46 and EBC-211, to facilitate enhanced keratinocytes wound healing responses. 4) Although differentially expressed genes of interest shown to be significantly different at the protein level are included in the main manuscript (by Western blotting and ImageJ densitometry), non-significant differences in Western blot protein profiles between untreated and epoxy-tigliane-treated keratinocytes are shown here.