Figure 3.

Effect of the Mutant BICD2 Alleles on the Golgi Apparatus

(A–P) HeLa cells were transfected with tGFP-BICD2 pCMV6-AC cDNA (either wild-type [A–C] or with BICD2 missense variants [D–P]). Cells were fixed and stained for a specific Golgi marker, Giantin (red) (1:100, Abcam), and DAPI (blue) (Life Technologies). Only transfected cells glowed green under the fluorescent microscope and allowed an easy comparison with untransfected cells, which reflected the wild-type situation. For better visibility, the Giantin staining alone is shown in the middle row and the inset in the right row, which represents an enlarged Golgi of a transfected cell. Scale bars represent 20 μm.

(A–C) The cells transfected with the wild-type construct show a condensed and abundant Golgi structure.

(D–P) Compared to wild-type or nontransfected cells, cells transfected with the three mutant cDNA constructs and the rare variant show a fragmented Golgi with a very diffuse and faint signal.

(Q) Quantification of cells with fragmented Golgi; 70–100 cells were counted for each experiment (n = 3) and are shown as the mean ± SD.