Fig. 1

ApoE4 increases cPLA2 but not iPLA2 expression in mouse primary astrocytes. Primary astrocytes were cultured from the cortex of P1-P4 mouse pups. A cPLA2 mRNA levels in primary astrocytes from ApoE3 and ApoE4-TR mice were measured by qPCR. B cPLA2, and phosphorylated cPLA2 (p-cPLA2) protein levels in primary astrocytes from ApoE3 and ApoE4-TR mice were detected by Western blot. C iPLA2 mRNA levels in primary astrocytes from ApoE3 and ApoE4-TR mice. D iPLA2 protein levels in primary astrocyte cultures from ApoE3 and ApoE4-TR mice was detected by Western blot. (n = 3 for each genotype for A-D). E, F Primary astrocytes from ApoE3 and ApoE4-TR mice were incubated with ³H-labelled AA (E) or ¹⁴C-labeled DHA (F) for 24 h, followed by induction by 100 nM ATP for 15 min. The efflux of ³H-AA (E) and ¹⁴C-DHA (F) from cells to media was measured by scintillation counting. G, H Primary astrocytes from C57BL/6 wild type mice were labeled with ³H-AA (G) or ¹⁴C-DHA (H) for 24 h, and then treated with rE3 or rE4 (0.2 μM) for 24 hours, followed by induction with 100 nM ATP for 15 min. ³H-AA (G) and ¹⁴C-DHA (H) efflux were measured by scintillation counting (n = 3 for E-H). Data are represented as mean ± SEM and analyzed by Student’s t-test (two-tailed). *p < 0.05, **p < 0.01. rE3: recombinant ApoE3, rE4: recombinant ApoE4