Fig. 5

Neuronal CLU facilitates neuron-to-astrocyte lipid transfer. A-B Lipid droplet (LD) staining by LipidTox in day 25 iGlut pure cultures (T/T vs. C/C). n = 6–7 coverslips per group (2 clones per line, 3–4 coverslips for each clone with 4–5 images per coverslip) from two independent differentiations of each line. Representative images in (A) from CD07 line. C-D LipidTox staining of Day 25 iGlut pure cultures after AAV-CLU overexpression. n = 5–6 coverslips per group (one clone from CD05 with genotype C/C, and 4–5 images of each coverslip) from two independent differentiations. Representative images in (C) from CD07 line. E The schematic diagram for the Red C12 lipid transfer assay. F-G Red C12 and LDs (BD493/503) signals in mAst. n = 7–8 coverslips per group (mAst co-cultured with 2 clones per line, 3–5 coverslips for each clone with 4–6 images from each coverslip) from two independent differentiations of each line. Representative images in (F) from CD05 line. H-I A Red C12 transfer assay was performed in the CLU overexpression system. iGlut were first infected with either AAV-eGFP or AAV-hCLU, as in (C). Red C12 was applied to the iGlut on Day 30, and its transfer to mAst was analyzed using the same procedures outlined in (E). n = 6 coverslips per group (mAst co-cultured with one clone of CD05 C/C line, and 6 coverslips for per group with 4–5 images from each coverslip) from two independent differentiations. J The schematic diagram for overexpression of CLU tagged with Flag. K Co-localization of neuron secreted and AAV-derived CLU (Flag +) and LDs (BD493/503 +) in mAst from day- 30 iGlut-mAst cocultures after AAV-hCLU infection (same experimental approach as Fig. 3G). Representative images from CD07 line. All statistical graphs depict mean ± SEM. Scale bars are indicated in corresponding image. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001