Tbl3 encodes a WD40 nucleolar protein with regulatory roles in ribosome biogenesis

Figure 3 Nucleolar localization of Tbl3-enhanced green fluorescent protein. NIH 3T3 cells were transfected with the p-enhanced green fluorescent protein (EGFP) N1-transducin β-like 3 (Tbl3) construct. Cells were examined by fluorescence microscopy 20 h. after transfection. A: Differential interference contrast (DIC) microscopy of three fibroblasts revealing the nucleoli; B: Merged tbl3-EGFP green fluorescence and DIC of the cells shown in panel A. The nucleolar localization of tbl3-EGFP was verified by confocal microscopy; C: DAPI staining of DNA in the nuclei of the same field; D: Green fluorescence alone of the same field; E: DAPI plus tbl3-EGFP; F: Western detection of the tbl3-EGFP fusion protein. NIH 3T3 cells expressing tbl3-EGFP were lysed in RIPA buffer. The lysate (5 μg protein) was run on a 4%-12% NuPAGE gel (lane 2) along with Magic Mark size markers (lane 1), blotted and probed with an anti-GFP monoclonal antibody and visualized by enhanced chemiluminescence.