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Fig. 1 | Molecular Neurodegeneration

Fig. 1

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

Fig. 1

Development of an AD model for identifying neuroprotective compounds. a Toxicity was determined by quantifying LDH release in the medium in BE (2)-M17 neuroblastoma cells incubated for 7 days with or without ATRA. The ATRA induced toxicity in the cells represents the “toxicity assay” used in all following experiments (n = 5; +/− ATRA: P < 0.0001; t = 7,346 DF = 8). b Cytosolic Ca2+ levels quantified by Ca2+-indicator Fura-2 fluorescence ratio (340 nm/380 nm) in hTAU-P301L cells after 6 days with and without ATRA treatment in the toxicity assay (n = 3; P = 0.0435; t = 2.915; DF = 4). c Genetic silencing of STIM1 reduces toxicity in the toxicity assay. # denotes the effect of STIM1 silencing compared to unsilenced cells, * indicates the effect of ATRA treatment (silencing: P < 0.0001; DF = 1; F (1, 36)=4048; ATRA treatment: P < 0.0001; DF = 1; F (1, 36)=8006; Corrected for multiple comparison using Tukey’s test: +/−ATRA - silencing: P < 0.0001; q = 132.4; DF = 36; −ATRA +/− silencing: P < 0.0001; q = 20.68; DF = 36; +/−ATRA + silencing: P < 0.0001; q = 46.53; DF = 36; +ATRA +/− silencing: P < 0.0001; q = 106.6; DF = 36). d Increased MAPK signalling prior to toxicity measured by western blot in the cells treated with or without ATRA (day 5). Data are normalised to the “control” (−ATRA) condition (n = 4; P = 0.0322, t = 2.775, DF = 6). A representative immunoblot is shown in the inset next to the graph. e The chemical structure of the REM compounds. R1 could be either Cl (REM0042826) or F (REM0043039) (Additional file 2: Supplementary Methods). f Concentration-response curves of ATRA-toxicity reducing effect REM0042826 (EC50 = 12 nM) and REM0043039 (EC50 = 5 nM) in the toxicity assay (n = 7 or 5 for REM0043039 or REM0042826 respectively). g Concentration-response curve (n = 3) of the cytosolic Ca2+ reducing effect of REM, (EC50 of 3 nM) in the toxicity assay. The dotted grey line represents the average basal Ca2+ levels of vehicle treated cells in the model (One-way ANOVA, REM treatment: P = 0.0387; F = 3.386; DF = 5)

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