Figure 3.

Missense mutations of GALC that interfere with secretion and cleavage are trapped in the ER. Representative confocal microscopy images of HeLa cells transiently transfected with either WT GALC (A), or the Krabbe mutations T513M and L618S (B). Cells were plated onto glass coverslips, fixed and immunostained using monoclonal antibody against GALC (green), the lysosomal marker cathepsin D (red) or the ER marker calreticulin (red). Nuclei were stained with DNA‐binding dye, DAPI (blue). Scale bar: 10 µm. C) To quantify colocalization, Pearson's correlation coefficients were calculated for each variant with both cathepsin D and calreticulin. Mean ± SEM for at least 20 individual cells from ≥3 independent experiments are shown. ***p ≤ 0.0001, calculated using a two‐tailed unpaired t‐test.