Fig. 2.

De novo protein synthesis of L5 and L11 is required for their accumulation in the ribosome-free form. (A) A549 cells were left untreated or treated with ActD for 5 h. Purified nucleoli were immunoblotted with the indicated antibodies. (B) H1299 cells containing YFP-L11 (green) were treated with ActD for 5 h, stained with antifibrillarin antibodies (red), and analyzed by CLSM. Fibrillarin nucleolar caps are indicated by arrowheads. Nuclei are outlined with dashed lines. (Scale bar, 5 μm.) (C) A549 cells were treated with ActD in the presence or absence of CHX for 1, 2, or 5 h, and nonribosomal fractions were immunoblotted with the indicated antibodies.