Supplementary Figure S4 from Intracellular Osteopontin Promotes the Release of TNFα by Mast Cells to Restrain Neuroendocrine Prostate Cancer

Supplementary Figure S4. Characterization of additional cell lines. A. Immunofluorescence for DAPI (blue) and CGA (red) on murine NEPC cells (TC566 and TC411K). B. Immunofluorescence for DAPI (blue) and OPN (red) on HMC-1 cells. C. Western blot for OPN in HMC-1 cells treated or not with Brefeldin A (BFA; 5 g/ml). Vinculin was evaluated as housekeeping control. D. Gating strategy applied to distinguish human NEPC cells (CD49f+c-Kit-) from HMC-1 MCs (CD49f-c-Kit+) by flow cytometry. E. OPN evaluation by ELISA (left panel) or real time PCR (the Spp1 transcript; right panel) in WT and OPN−/− MCs, and in MC/9, T23, and ST4787 cells. F. Elisa for OPN in parental MC/9 cells and in MC/9-CTR, MC/9-OPNf, MC/9-iOPN cells. T23 cells were used as control.