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. 2006 May;17(5):2287–2302. doi: 10.1091/mbc.E05-11-1089

Figure 1.

Figure 1.

hRad21 and NIPBL knockdown cells are mitotically delayed without securin destruction. (A) Time-lapse images taken from movies (Supplement 2) are shown. Top, control no RNAi; middle, NIPBL RNAi (120 h after RNAi); bottom, hRad21 RNAi (104 h after RNAi). Arrow, the partially formed plate. (B) The duration from NEBD until the onset of anaphase was measured in the movies of control non-RNAi, NIPBL, and hRad21 RNAi cells. (C) hRad21 retention in nuclei. Transfected cells were extracted in CSK buffer supplemented with Triton X-100 before fixation. Cells shown are 96 h after transfection. Top, DNA stained with Hoechst 33342; bottom, immunostaining of hRad21. (D) Top, left, normal metaphase; middle, cell showing a few misaligned chromosomes (arrowhead); right, cell showing abundant misaligned chromosomes. Bottom, the frequencies of each appearance measured in control non-RNAi and hRad21 RNAi cells. (E and F) Securin and cyclin B1 were immunostained in control non-RNAi and hRad21 RNAi cells fixed by paraformaldehyde. In hRad21 RNAi cells, mitotic cells containing abundant EMCs are shown. Bars, 10 μm.