Fig. 6

Progranulin Interacts with GCase. HEK-293 cells were co-transfected with constructs expressing HA-tagged human progranulin and/or myc-flag-tagged human GCase. HA-tagged progranulin was then immunoprecipitated from cell lysates with an anti-HA antibody. a, Flag-tagged GCase co-immunoprecipitated with progranulin, indicating interaction of the two proteins. b, Consistent with the co-immunoprecipitation of GCase with progranulin, we detected strong proximity ligation (PLA) signal in HEK-293 cells co-transfected with human progranulin and human GCase constructs. c, The specificity of the Flag-HA PLA signal was confirmed by the presence of significantly more PLA puncta from cells co-transfected with the progranulin-HA and GCase-Flag constructs than in cells transfected with only one of the constructs, or from cells that underwent PLA in the absence of the HA and Flag antibodies (ANOVA effect of experimental condition, p < 0.0001, *** = p < 0.001 and **** = p < 0.0001 by Dunnett’s post-hoc test). The scale bars represents 5 μm. GCase = β-glucocerebrosidase, GRN = progranulin, PLA = proximity ligation assay