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Fig. 1. The number and size of GWBs varies during the cell cycle. (A-C) HEp-2 cells were co-stained with the index human anti-GW182 serum (A, green; nuclei counterstained with DAPI, blue) and rabbit anti-CENP-F antibody (B, red). CENP-F is known to be expressed predominantly in cells in late S or G2 phase and throughout mitosis (B, arrows). (C) The merged image of GW182 and CENP-F. GWBs show cell-cycle-dependent expression, with the largest bodies (arrowheads) detected primarily within cells in the late S or G2 phase. The number of GWBs also is higher in cells with the strongest expression of CENP-F. Staining of GW182 in cells at anaphase (a), telophase (t), and G1 stages are indicated by double-arrows (A). GWBs are sometimes observed adjacent to the nuclear membrane (thin arrowheads). Scale bar: 10 µm. (D) Western blot analysis of the index human anti-GW182 serum (lane 1) and a control normal human serum (lane 2) using HEp-2 cell lysates, separated by SDS-PAGE using a 12.5% gel and transferred to a nitrocellulose membrane.

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