Journal: Cellular Logistics

Article Title: Centrosomal AKAP350 modulates the G 1 /S transition

doi: 10.4161/cl.26331

Figure Lengend Snippet: Figure 2. Characterization of AKAP350CTD cells. AKAP350CTD1 and AKAP350CTD4 cells were cultured for 24 h, fixed with methanol and stained for γ-tubulin ( A ) or fixed with paraformaldehyde and stained for AKAP350 ( B ). Immunofluorescence images were obtained by confocal microscopy. ( A ) Images show γ-tubulin staining and AKAP350CTD-GFP fluorescence. Arrows indicate the centrosomes with orthogonal view. ( B ) Images show the z-projection of AKAP350 staining. Arrows indicate centrosomal localization of AKAP350. Bars represent the mean ± s.e.m. of the % of AKAP350 fluorescence intensity associated with centrosomes for three independent experiments. Bar: 5 μm.

Article Snippet: Fluorescence localization was detected by confocal laser microscopy (Nikon C1SiR with inverted microscope Nikon TE200).

Techniques: Cell Culture, Staining, Immunofluorescence, Confocal Microscopy, Fluorescence

Journal: Cellular Logistics

Article Title: Centrosomal AKAP350 modulates the G 1 /S transition

doi: 10.4161/cl.26331

Figure Lengend Snippet: Figure 5. Effect of AKAP350CTD expression on centrosomal levels of Cdk2. ( A ) AKAP350CTD and control cells were cultured for 24 h and fixed with methanol. Fixed cells were double-stained for Cdk2 and γ-tubulin, and analyzed by confocal microscopy. The first and second column show images of the channels corresponding to γ-tubulin and Cdk2, and the third column the overlay view in RGB mode of the channels corresponding to GFP expression (green) and γ-tubulin (blue) and Cdk2 (red) staining. Bar, 10 μm. Arrows indicate centrosomes with orthogonal views. ( B ) Centrosomal enriched fractions (CF) were prepared as described in Materials and Methods, and Cdk2 levels at CF and at the starting material (SM) analyzed by western blot. γ-tubulin was used as loading control. Bars show the percentage of Cdk2 recovered at CF. Results are mean ± s.e.m of 4 independent experiments. * P < 0.05.

Article Snippet: Fluorescence localization was detected by confocal laser microscopy (Nikon C1SiR with inverted microscope Nikon TE200).

Techniques: Expressing, Control, Cell Culture, Staining, Confocal Microscopy, Western Blot