Additional file 1: of Modifying Rap1-signalling by targeting Pde6δ is neuroprotective in models of Alzheimer’s disease

Figure S1. ATRA induced cell death in the model is enhanced by expression of a cDNA encoding mutant human Tau and is prevented by REM. a Left: Western blot analysis confirmed successful tau overexpression (approximately 2-fold) in neuroblastoma cells containing the hTAU-P301L plasmid. Right: Toxicity was determined by quantifying LDH release in the medium in BE (2)-M17 neuroblastoma cells with (hTAU-P301L) and without (Control) expression of a cDNA encoding hTAUP301L incubated for 7 days with or without ATRA. The ATRA induced toxicity in hTAU-P301L cells represents the “toxicity assay” used in all following experiments (two-way ANOVA: +/− ATRA: P < 0.0001, F(1, 36)=4276, DF = 1; +/− hTAU-P301L: P < 0.0001, F (1, 36)=255.3, DF = 1; Sidak’s multiple comparison test: control +/− ATRA: P < 0.0001, t = 27.54, DF = 36; hTAU-P301L +/− ATRA: P < 0.0001, t = 64.93, DF = 36; n = 10). # denotes the effect of hTAU-P301L expression silencing compared to control cells, * indicates the effect of ATRA treatment. b Representative images of neuroblastoma cells exposed to ATRA for six days treated with either REM or vehicle. c Treating hTAU-P301L cells in the toxicity assay with tau aggregation inhibitior methylthioninium rescues cell death ((EC50 = 23 nM; n = 3) One-way ANOVA, Methylthioninium treatment: P < 0.0001, F = 109.7, DF = 4). (TIF 1286 kb