Fig. S3

DFO does not prevent cell death in SH-SY5Y cells expressing TRPM7-DN, whether or not cells are induced to differentiate with retinoic acid. A) SH-SY5Y cells were plated in 12-well plates at a density of 4 x 104 cells/well in complete media. An adenovirus expressing the TRPM7-E1047K pore mutant (TRPM7-DN) at a multiplicity of infection (M.O.I.) of 30 was used to suppress TRPM7 channel function. To assess whether DFO is protective against TRPM7-DN-indcued cell death, the drug was co-administered at increasing concentrations (μM). Cells treated with DFO alone exhibited cell death. Non-transduced (NT) and cells transduced with an adenovirus expressing LacZ were used as negative controls. Cells were counted 72 hrs after plating and are expressed as a percentage of non-transduced cells (% NT). An adenovirus expressing GFP (AdGFP) was used to assess viral transduction efficiency at an M.O.I. of 30. Shown are relief contrast (RC) and fluorescence (FL) images of virally transduced cells. Scale bar, 200 μm. B) SH-SY5Y cells were plated in 12-well plates at a density of 4 x 104 cells/well in complete medium. SH-SY5Y cells were induced to differentiate by culturing in complete medium with 10 μM retinoic acid (RA) for 72 hrs. After 72 hrs of differentiation, cells were transduced with TRPM7-DN at a M.O.I. of 30, and DFO was co-administered at increasing concentrations (μM). Cells treated with DFO alone exhibited cell death. Non-transduced cells and cells transduced with an adenovirus expressing LacZ were used as negative controls. Cells were counted 6 days after plating.