Figure 1. Expansion of the Golgi following Scyl1 knock down.

A, HeLa cells were transfected with control (ctrl) siRNA or one of two previously characterized siRNAs (Scyl1 #1 and #2) specific for Scyl1 [16]. The cells were subsequently fixed and processed for indirect immunofluorescence with antibodies against Scyl1 (red) and either GM130 or 58K (green). Magnified views of regions of the GM130 staining are shown in the rightmost 3 panels. The scale bars = 10 µm. B, The two-dimensional surface area was calculated for a large number of Golgi as described under Materials and Methods. Despite variability in Golgi size as indicated by the overlapping error bars, which indicate standard error of the mean, Scyl1 knock down causes a significant increase in the surface area of the Golgi (*** p<0.001) as determined by a Student's t-test. C, Cells treated with ctrl siRNA or Scyl1 #1 siRNA were subsequently mixed and co-stained for Scyl1 (red) and 58K (green). The two cells in the middle, lacking Scyl1 expression have expanded Golgis. The scale bar = 10 µm. D, HeLa cells, transfected with control siRNA or Scyl1 #1 siRNA were subsequently fixed and processed for indirect immunofluorescence with antibodies against Scyl1 (red) and GM130 (green). Outlines of the cells are indicated. The scale bar = 10 µm. E, The two-dimensional surface area of the Golgi was calculated and presented as a percentage of the two-dimensional whole cell area. The Golgi in Scyl1 knock down cells covers a significantly (*** p<0.001, Student's t-test) larger area of the cell. Error bars represent standard error of the mean for all graphs.