Fig. 3

GCS inhibition by GENZ increases IR on the surface of mHippoE-14 neurons exposed to ADDLs. a Quantitative PCR shows that IR mRNA is not changed by GCS inhibition. b A surface biotinylation assay and subsequent western blot analysis of the precipitated IR show GENZ-dependent sequestering of the IR to the cell surface (n = 5). c A PLA using two different IR antibodies depicts surface IR levels on non-permeabilized cells. GENZ treatment increases surface IR levels (n = 97–135 cells). d Immune fluorescence shows surface IR of non-permeabilized cells. ADDL exposure decreases surface IR levels (white bar). However, surface IR levels are increased in cells treated with GENZ (grey bar; n = 121–212 cells). e A PLA on non-permeabilized cells using two IR antibodies (N-20 and D-17) confirms that GENZ treatment increases surface IR upon ADDL exposure (n = 119–186 cells). Unpaired two-tailed student’s t-test (if p ≤ 0.05, or p ≤ 0.001 results are marked with (*) or (***), respectively); 5 μM ADDLs, 24 h. Means ± SEM. Scale bars: 10 μm