Fig. 5

FIG4 KO fibroblasts display exaggerated swollen compartments. a Control fibroblasts were depleted for FIG4 by CRISPR/Cas9 and mixed populations were grown. Western blotting confirms loss of FIG4 protein. b FIG4 KO fibroblasts display swollen LAMP1 positive compartments that can also be seen by Differential Interference Contrast (DIC). c FIG4 KO fibroblasts contain vacuolar compartments visualised by conventional electron microscopy. d Correlative Light and Electron Microscopy (CLEM) of FIG4 KO fibroblasts transfected with the PtdIns(3)P probe, PX-GFP. GFP fluorescence displays overlap with smaller endocytic organelles, but not large compartments