Immunolocalization of SYT2. Haiyan Zhang Liang Zhang Bin Gao Hai Fan Jingbo Jin Miguel A. Botella Liwen Jiang Jinxing Lin 10.1371/journal.pone.0026477.g004 https://plos.figshare.com/articles/figure/_Immunolocalization_of_SYT2_/379846 <p>Bars = 10 µm. (A) Protein gel blot of SYT2 in wild-type (WT), <i>syt2-1</i>, and SYT2-GFP overexpressing plants (SYT2-GFP1 and SYT2-GFP2 are different lines). The blots were probed with polyclonal anti-SYT2 (Right) and monoclonal anti-GFP (Left) antibodies to detect the SYT2 and GFP-tagged proteins, respectively. The expected sizes of the proteins are indicated. The bottom images show Coomassie Brilliant Blue staining (CBB) as loading controls. (B) RT-PCR analysis of <i>SYT1</i> and <i>SYT2</i> in <i>syt2-1</i> and wild-type plants. <i>Actin</i> served as a control. (C and D) Localization of SYT2 in root cells of wild-type plants. Root tissues of <i>Arabidopsis</i> grown on ½ MS solid medium for 3–4 days were prepared for immunolabeling with normal rabbit serum (as a control) (C), or anti-SYT2 antibody as the primary antibody (D) and fluorescein isothiocyanate (FITC)-labeled anti-rabbit IgG as the secondary antibody. (E) Double-labeling with anti-SYT2 and anti-GFP antibodies in root cells of SYT2-GFP-overexpressing seedlings. Anti-SYT2 and anti-GFP antibodies were labeled with tetramethylrhodamine-5-isothiocyanate (TRITC)-labeled anti-rabbit IgG and FITC-labeled anti-rat IgG, respectively. Arrows indicate the overlap of green and red fluorescent signals. (F) Double-labeling with anti-SYT2 and anti-GFP antibodies in root cells containing Golgi marker ST-YFP. Anti-SYT2 and anti-GFP antibodies were used as in (E). Arrows indicate the overlap of green and red fluorescence signals. (G–L) Immuno-gold labeling and electron microscopic observation showed that SYT2 was located on Golgi apparatus in root tip cells of <i>Arabidopsis</i>. (G and H) Electron microscopic observation showed that SYT2 was located mainly on Golgi apparatus in root tip cells of wild type plants. (H) High-magnification image of Golgi apparatus in the inset in (G). (I and J) Immuno-gold labeling of Golgi apparatus in root tip cells of <i>SYT2</i>-overexpressing plants. (J) High-magnification image of Golgi apparatus in the inset in (I). (K and L) Control section, incubated with the secondary antibody alone, did not show gold particles on Golgi apparatus. G: Golgi apparatus. Bars: 2 µm (G, I,); 50 nm (H, J, K, L).</p> 2011-11-28 02:44:06 plant biology cell biology