Figure S1. Enzastaurin reduced expression levels of phospho-PKC substrates and increases doxorubicin-induced apoptosis in SAN melanoma.

Figure S2. (A) Enzastaurin decreases melanoma growth in a dose–response manner. After a 72 h culture, cell growth was measured by MTS assay (CellTiter 96 AQueous One Solution Cell Proliferation Assay; Promega, WI, USA), according to the manufacturer's protocol. Metabolically active SAN cells were measured by adding 20 μl of MTS tetrazolium to each well. After 2 h of incubation, the plates were analysed using 490 nm absorbance, with a Multilabel Counter (BioTek, Milan, Italy). (B) Dose–response to doxorubicin. Values represent means and standard deviations of 12 experiments (3 for each cell lines, in triplicated) performed with G361, SK-MEL3, A375 and SAN. Cell death was measured after a 48 h incubation with flow cytometry and propidium iodide incorporation.

Figure S3. An electrophoretic mobility shift assay (EMSA) using nuclear extracts prepared from Jurkat cells stimulated with 20 ng/ml TNFα for 1 h, in the absence or the presence of 100 ng/ml H398 antibody.

Figure S4. PKCζ overexpression counteracts enzastaurin-induced increase in doxorubicin-induced cell death.

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