Figure 6.

Altered glucose sensing contributes to the hyperactive autophagosome formation in Synj1-deficient astrocytes.A, representative images of cultured astrocytes from P0 synj1 WT, HET, and KO brains expressing GFP and immunolabeled with anti-GluT-1. B, box plots for membrane immunofluorescence of GluT-1 in cultured astrocytes. Data from three independent cultures. p Values were from Tukey's post hoc tests following one-way ANOVA. C and D, representative Western blots for WT and KO astrocytes at the basal condition (ctrl) as well as those that treated with rapamycin (Rap) or serum starvation (DMEM) for 4 h (C). Analysis for AMPK activity and expression in four independent cultures at the basal level. p Values are from Student's t test. E, representative images of cultured astrocytes from P0 synj1 WT and KO brains sparsely expressing GFP-LC3 or GFP-LC3 with FLAG-GLUT1. Cells were fixed and immunolabeled with anti-GFP and FLAG. White box indicates the contrast-adjusted GFP-LC3 signal in the GLUT1 expressing cell. F, box plots for the number of GFP-LC3 puncta in WT, KO, and KO + GLUT1 astrocytes. p Values are from Tukey's post hoc tests following one-way ANOVA. AMPK, AMP-activated protein kinase; DMEM, Dulbecco's modified Eagle's medium; HET, heterozygous; Synj1, Synaptojanin1.