Figure 4.

Synj1 deficiency impairs autophagy clearance.A, representative images of cultured astrocytes from P0 synj1 WT, HET, and KO brains expressing GFP-LC3 (ctrl), those that were treated with 200 nM rapamycin (Rap + baf) or the DMEM-only medium (DMEM + baf) for 4 h including a 20 nm bafilomycin A1 treatment in the last hour. WT and HET ctrl images were reused from Figure 2A. B, box plots comparing the number of LC3 puncta between the ctrl and the rapamycin-treated groups (left) or between ctrl and the DMEM-only medium treated groups (right). p Values were from Mann–Whitney U tests. C, representative images of cultured astrocytes from P0 synj1 WT and KO brains expressing GFP-LC3 (ctrl), those that were treated with 200 nM rapamycin for 4 h (Rap), and those that were treated with the DMEM-only medium for 4 h. Cells were fixed simultaneously and immunolabeled with GFP and p62. D, box plots comparing the normalized p62 levels across in the ctrl and the rapamycin-treated groups (left) or between ctrl and the DMEM-only medium treated groups (right). p Values are from Tukey's post hoc test following two-way ANOVA. Data from four independent batches of cell cultures. DMEM, Dulbecco's modified Eagle's medium; HET, heterozygous; Synj1, Synaptojanin1.