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However, how they regulate specific processes during meiosis is not well understood. = aggregates.Įpigenetic modifiers are emerging as important regulators of the genome. (C) Table shows the percentage of -1 oocytes at diakinesis displaying each one of the indicated defects in chromosome morphology. TZ, transition zone EP, early pachytene MP, mid-pachytene LP, late pachytene.
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***P<0.0003 by the two-tailed Mann-Whitney test, 95% C.I., after Bonferroni correction. Error bars represent SEM for technical repeats from two biological replicates. A significant decrease in levels of RAD-51 foci were observed for zones 3 to 5 in zfp-1 germlines compared to wild type and for zones 3 to 7 in zfp-1 syp-1 germlines compared to syp-1.
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Between 4 and 6 gonads were scored per genotype. (B) Histogram shows the mean number of RAD-51 foci/nucleus (y-axis) scored along each zone in the germlines (x-axis) of the indicated genotypes. (A) High-resolution images representative of mid-pachytene nuclei (zone 5) immunostained for RAD-51 (magenta) and co-stained with DAPI (blue). S4 Fig: zfp-1 mutant worms show altered DSB formation and bivalent morphology defects. 10% (1/10) of these nuclei exhibited discontinuities in HTP-3 signal and in 70% (7/10) the unsynapsed chromosome corresponded to the X chromosome as determined by presence of HIM-8 signal (arrowhead). Unlike wild-type worms, 5.78% of nuclei (10/173) in zfp-1 mutants exhibited DAPI-stained regions lacking SYP-1 signal at the pachytene stage (arrow). Right, high-resolution images of mid-pachytene nuclei co-immunostained with SYP-1 (green), HTP-3 (red), HIM-8 (yellow) and DAPI (blue). 4 gonads from two biological repeats were scored for wild type (n = 154) and zfp-1 (n = 173) n = number of nuclei scored. Nuclei showing complete overlapping signal of the lateral element component HTP-3 and the central region component SYP-1 along all chromosomes were considered as nuclei with complete synapsis. (C) Left, histogram indicating the percentage of nuclei that exhibit complete synapsis as a function of meiotic progression in wild-type and zfp-1 gonads.
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Chromosomes were scored as paired when two signals were ≤ 0.75 μm apart from each other. Right bottom, histogram representing the percentage of nuclei with paired FISH signal (5S rDNA) scored at different zones along the germline in wild type and zfp-1. Yellow arrow indicates nucleus with unpaired FISH signals. Top right, high-resolution images of early pachytene nuclei (zone 4) stained with DAPI and hybridized with a FISH probe against the 5S rDNA locus located near the center of chromosome V (green). X chromosomes were scored as paired when the two HIM-8 signals were ≤ 0.75 μm apart from each other. Bottom left, histogram representing the percentage of nuclei with paired HIM-8 signals scored at different zones along the germline in wild-type and zfp-1 worms. Yellow arrow indicates nucleus with unpaired HIM-8 signal. (B) Top left, high-resolution images of early pachytene nuclei (zone 4) co-stained with HIM-8 (magenta) and DAPI (blue). At least 30 gonads from two independent biological repeats were analyzed for each genotype. Hatched lines demarcate the first (left) and the last (right) rows with nuclei showing SUN-1 (pS8) signal. Asterisks indicate the premeiotic tip and white arrows show the direction in which nuclei move proximally in the germline (meiotic progression). zfp-1 mutant worms showed an extension in SUN-1-positive signal compared to wild type suggesting defects in meiotic progression. (A) Whole mounted gonads of wild-type and zfp-1 worms co-immunostained with SUN-1(pS8) (green) and DAPI (blue). S3 Fig: zfp-1 mutant worms exhibit defects in meiotic progression, delayed pairing and problems with SYP-1 loading.