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Fig. 3. The replication band of the macronucleus (rb, replication band; fz, forward zone; rz, rear zone). (a-c) Mid optical section of a macronucleus. a and b, Fluorochrome channels were scanned sequentially generating 8-bit greyscale images; a, To-Pro-3; b, BrdU pulse-labelling of replication foci (pulse 45 minutes) c, Overlay, with false colours assigned to each channel (To-Pro-3, blue; BrdU, red). (d) Average intensity projections (XY plane, YZ plane) of 63 serial optical sections. 3D reconstruction of the nuclear surface (blue) and the disc-shaped replication band (red). The surface was partly cut to look inside. (e) Sequential labelling of replication sites with IdU and CldU (pulse 30 minutes each, chase 5 minutes). Fluorochrome channels were scanned sequentially generating 8-bit greyscale images. Overlay with false colours assigned to each channel (IdU, green; CldU, red). Nuclear surface was rendered using To-Pro-3 counterstaining and marked by a white line in the overlay image. Sites of simultaneous IdU and CldU incorporation appear yellow. Replication occurs in hundreds of replication foci with a size of approximately 0.5 µm each. Initially, replication foci are more concentrated in the periphery (red). Later initiation of replication foci extends to the interior (yellow and green).
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