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Fig. 4 | Molecular Neurodegeneration

Fig. 4

From: Sex-dependent calcium hyperactivity due to lysosomal-related dysfunction in astrocytes from APOE4 versus APOE3 gene targeted replacement mice

Fig. 4

Dysregulated Ca2+ excitability in APOE4 immortalized astrocytes is due to greater lysosomal V-ATPase activity. a Schematic of the lysosomal Ca2+ mobilization and uptake mechanisms. b Representative traces and quantification of ATP-induced cytosolic Ca2+ responses of cells treated with 0.1% DMSO or 100 μM Ned-19 (diluted in 0.1% DMSO) for 20 min (N = 3). c Quantification by real time qPCR of mRNA expression of Tpc1 and Tpc2 (normalized to Gapdh mRNA contents, N = 5) and Trpml (normalized to Gapdh mRNA contents, N = 4). d Representative traces and quantification of the area under the curve (AUC) of intracellular Ca2+ after lysing the lysosomes with 200 μM GPN (N = 4). e Representative traces and quantification of ATP-induced Ca2+ responses after treating cells with DMSO or 2 μM bafilomycin A1 (Baf) for 20 min (N = 3). Note that the black arrow indicates differences in basal Ca2+. f Quantification of V-ATPase subunit V0D1 with western blot, normalized to β-actin expression (N = 4). g Quantification of the pH of acidic organelles in APOE3 and APOE4 astrocytes (N = 3). Unpaired parametric T-test was used, except in b and e, where one-way ANOVA was used. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***)

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