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. 2012 Jan 20;109(6):1979–1984. doi: 10.1073/pnas.1108705109

Fig. 3.

Fig. 3.

The kinetochore is transcriptionally active during mitosis. (A) In situ transcription assay revealed transcriptional activity in cytospun NIH 3T3 interphase cells, as shown by nuclear and nucleolar FITC-rUTP incorporation (arrow). (B) The in situ transcription assay showed FITC-rUTP incorporation at the kinetochore regions (arrows) of the mitotic chromosomes. (C) FITC-rUTP incorporation at the kinetochore is sensitive to the RNAPII inhibitor α-amanitin. (D) CREST immunofluorescence performed after the in situ transcription assay showed that the FITC-rUTP signals colocalized with CREST signals (arrows). (E) Fluorescence intensity line scans of a single chromosome (boxed in D). Lateral (solid line) and axial (dashed line) fluorescence intensity line-scans through the kinetochores confirmed that the FITC-rUTP signals (green lines) colocalized with CREST signals (red lines); blue lines indicate DAPI staining. (Scale bars: 5 μm in A–D, 2 μm in E.)