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. 2005 May;16(5):2395–2413. doi: 10.1091/mbc.E04-11-0992

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Figure 9. — Formation of nucleolar caps is a dynamic energy-requiring process. Immunofluorescence microscopy of (A) The formation of PSF concave nucleolar caps at different time points after addition of ActD. Hoechst DNA counterstain is seen in blue. (B) Western blot analysis of PSF from protein extracts of cells at time points as in A. (C) Recovery curves after photobleaching of GFP-PSF, GFP-fibrillarin, GFP-Nopp140, and GFP-p14(ARF) in the nucleoplasm or nucleolus before ActD treatment (green) and in nucleolar caps after ActD treatment (orange). (D) PSF and nucleolin labeling of cells treated with: (a) ActD only, (b) ActD plus Na-azide and 2-deoxyglucose. (E) Formation of PSF concave caps. ActD treatment was performed under different conditions: (a) HBSS medium, (b) HBSS medium plus 10% FCS, (c) HBSS medium with glucose, (d) HBSS medium with pyruvate, (e) DMEM without FCS. Bar, 10 μm.