JCS03054 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. SYN1 immunostaining in wild-type and afd1 nuclei at pachytene. In the wild type, a bright SYN1 signal (green in merged images) is observed between homologous chromosomes (DAPI in red in merged images). In afd1, we enhanced the signal to detect any signal on the chromosomes, thereby increasing the background signal. No staining was detected on the afd1-1 and afd1-2 chromosomes. In afd1-3, patches of signal were detected in the nucleus (arrows). In afd1-4, the AFD1 signal was diffuse and colocalized with the chromosomes. Individual channels are shown in black and white for clarity. Bar, 5 μm.

• Supplemental Figure 2 -

  Fig. S2. AFD1 and ASY1 immunostaining (4% paraformaldehyde fixation, whole nuclei shown). At pre-meiotic interphase, a diffuse AFD1 (blue) and ASY1 (green) signal is detected in wild-type nuclei. At zygotene in unsynapsed regions, the AFD1 and ASY1 signals colocalized. However, in synapsed regions the ASY/HOP1 signal is less intense, whereas the AFD1 signal is brighter. At pachytene, chromosomes are completely synapsed except at the nucleolar organizing region (NOR). AFD1 staining covers the whole chromosomes whereas ASY1 staining only localizes to the NOR (arrowhead). Bar, 5 μm.

• Supplemental Figure 3 -

  Fig. S3. ImmunoFISH analysis of telomeres and AEs. (A) ASY1 (blue) is recruited on the entire chromosomes (red) and there is no preferential localization of ASY1 near the telomeres (green) in the wild type. Individual channels are shown in black and white for clarity. (B) AFD1 (blue) is also recruited along the entire length of the chromosomes (red) and not preferentially near the telomeres as shown on a median section of a wild-type nucleus. In afd1-4, we did not detect a stronger AFD1 signal (blue) near the cluster of telomeres (green). Bar, 5 μm.

• Supplemental Figure 4 -

  Fig. S4. RAD51 immunostaining in afd1-3 and afd1-4: serial sections. Most of the RAD51 foci (green) are forming long aggregates (green) in both afd1-3 (A-D) and afd1-4 (E-H). Four sequential 0.3 μm optical sections are shown. Bar, 5 μm.