Colocalization of SMN with the Golgi apparatus.

<p>(A–C) NSC34D cells were stained with SMN antibody (green) and antibodies against Gm130 (A), Tgn38 (B), and Golgin-97 (C) (red). Magnified regions are shown in boxes at right of the merged figures. Obvious colocalization between each protein is indicated with pink arrows. Scale bar: 20 µm; 2 µm in magnified box. (D) In the control group, antibodies against mouse (Ms) or rabbit (Rb) immunoglobulin G (IgG) were used in place of SMN antibody or the Golgi marker antibodies. (E) Quantitation of the colocalization ratio between SMN with Gm130, Tgn38, Golgin-97, and WGA in selected Golgi enriched region. Data represent the mean ± SEM from three independent experiments (over 50 cells were counted in each group). ***, <i>P<</i>0.001. <i>ns</i>: not significant. (F) EGFP-SMN and dsRed-β1,4-Gal-T or (I) EGFP-SMN and mCherry-FYVE are shown in green and red, respectively. Time series images were taken every 10 sec, including <i>z</i>-stacks. Images are shown in three dimensions with views from the front, underside, and side-back (F). Selected planes 36–43 (F) and 1–8 (I) are shown. Arrows indicate obvious colocalizations of SMN and the Golgi. (G, J) Transfection efficiencies of EGFP-SMN with dsRed-β1,4-Gal-T were 49.4% ±1.0% with 41.1% ±1.2%, respectively; efficiencies for EGFP-SMN with mCherry-FYVE were 47.5% ±2.3% with 43.8% ±2.2%, respectively. Data represent the mean ± SEM, <i>n</i> = 4 in each group. (H, K) Quantitation of the colocalization ratio in each time-point between SMN with organelle markers in a selected view from a neurite. Bar: 2 µm.