Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Mapping of two centromeric sequences, CentA and CentC, on pachytene chromosomes of KYS. (A) and (B) Pachytene chromosomes were simultaneously probed with the CentA element (green signals in A) and CentC repeat (red signals in B). (C) The merged image shows the colocalization of two centromeric sequences. Bar = 10 μm.
• Supplemental Figure 2 - Mapping of 22 kD alpha-zein cluster on pachytene chromosome 4 using HR gene and 3-D FISH. (A) Pachytene chromosomes derived from squash preparations were probed with the FITC-labeled zein gene and Cy3-labeled CentC and telomere sequences. The magnified image of signals is shown in the inset. The computationally straightened chromosome 4 taken from (A) is shown in (B). (C) A projection after deconvolution of a 3-D pachytene meiocyte shows signals of zein (green), centromere (red) and telomere (red). The magnified image of signals is shown in the inset. The computationally straightened chromosome 4 taken from (C) is shown in (D). Two straightened chromosomes are shown at the same magnification. Bar = 10 μm.
• Supplemental Figure 3 - The effect of pepsin treatment on maize KYS pachytene chromosomes. The chromosomes were prepared by the squash procedure as described in methods. After RNase digestion, the chromosomes were treated with 1 μg/ml pepsin at 37°C, followed by washes in water as described in methods. After dehydration in ethanol chromosomes were observed by Nomarski microscopy and images were captured using a CCD camera. (A) The pachytene chromosomes before pepsin treatment display cellular debris uniformly covering the chromosomes. Under this condition, there is usually some background staining covering chromosomes after FISH. (B) The pachytene chromosomes after 10 min in pepsin were still covered by debris. (C) The same cell as shown in (A) after a 30 min treatment with pepsin. This example shows chromosomes at a stage suitable for FISH mapping. (D) Pachytene chromosomes after 60 min treatment of pepsin have a degraded chromosome morphology that is unsuitable for FISH. Bar = 10 μm.
• Supplemental Table 1
• Supplemental Table 2