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Gene Expression Data
Assay Details
Assay
Reference: J:275851 Harris C, et al., Conversion of random X-inactivation to imprinted X-inactivation by maternal PRC2. Elife. 2019 Apr 2;8:e44258
Assay type: Immunohistochemistry
MGI Accession ID: MGI:7495828
Gene symbol: Eed
Gene name: embryonic ectoderm development
Antibody: anti-EED
Detection system: Secondary antibody coupled to Alexa Fluor 555
Results
Specimen 1A female: embryonic day 3.5; Tg(CAG-EGFP)D4Nagy/0 (more )
Note: Tg(CAG-EGFP)D4Nagy was used to sex the embryo.
Structure Level Pattern Image Note
TS4: blastocyst Present 1A EED female , 1A lower EED female Accumulation of Eed, H3K27me3 with Xist RNA in a vast majority of the nuclei (72-100%).
Specimen 1A male: embryonic day 3.5 (more )
Note: Tg(CAG-EGFP)D4Nagy was used to sex the embryo.
Structure Level Pattern Image Note
TS4: blastocyst Absent 1A EED male , 1A lower EED male
Specimen 1B +/-: embryonic day 3.0-3.5; Eedtm1d(EUCOMM)Wtsi/Eed+ (more )
Note: Tg(Prm-cre)58Og was used to delete the Eed floxed allele in the male germline.
Structure Level Pattern Image Note
TS4: blastocyst Present 1B lower EED +/- , 1B EED +/- Coincident accumulation of Eed and/or H3K27me3 with Xist RNA in over 70% of the nuclei (9 out of 41 female embryos), similar as in the wild-type female.
Specimen 1B -/-: embryonic day 3.0-3.5; Eedtm1d(EUCOMM)Wtsi/Eedtm1d(EUCOMM)Wtsi (more )
Note: Tg(Prm-cre)58Og was used to delete the Eed floxed allele in the male germline.
Structure Level Pattern Image Note
TS4: blastocyst Absent 1B EED -/- , 1B lower EED -/- Devoid of Eed or H3K27me3 enrichment overlapping with the Xist RNA coat (9 out of 41 female embryos).
Specimen not shown: embryonic day 3.0-3.5; Eedtm1d(EUCOMM)Wtsi/? (more )
Note: Tg(Prm-cre)58Og was used to delete the Eed floxed allele in the male germline.
Structure Level Pattern Image Note
TS4: blastocyst Present 23 out of 41 female embryos displayed 2-70% of nuclei with Eed and/or H3K27me3 enrichment, likely representing Eed+/- or Eed-/- embryos that had not yet fully depleted maternally-inherited Eed protein or Eed+/- embryos which had not yet robustly expressed zygotic Eed.
Specimen not shown: embryonic day 3.0-3.5; Eedtm1d(EUCOMM)Wtsi/? (more )
Note: Tg(Prm-cre)58Og was used to delete the Eed floxed allele in the male germline.
Structure Level Pattern Image Note
TS4: blastocyst Absent No enrichment of Eed or H3K27me3 in the nucleus, as in the WT male embryos.
Specimen 2A 2-cell fl/fl: embryonic day 1.0; Eedtm1c(EUCOMM)Wtsi/Eedtm1c(EUCOMM)Wtsi (more )
Note: Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS2: 2-cell stage embryo Present 2A fl/fl
Specimen 2A 2-cell fl/- or -/-: embryonic day 1.0; Eedtm1d(EUCOMM)Wtsi/? (more )
Note: Tg(Prm-cre)58Og was used to delete the Eed floxed allele in the male germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS2: 2-cell stage embryo Present 2A fl/- or -/- Exhibited similar levels of Eed and H3K27me3 as in the control (summarized in 2C and 2E).
Specimen 2A 2-cell m-/-: embryonic day 1.0; Eedtm1d(EUCOMM)Wtsi/Eed+ (more )
Note: Tg(Zp3-cre)93Knw was used to delete the Eed floxed allele in the female germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS2: 2-cell stage embryo Absent 2A m-/- Devoid of both Eed and H3K27me3 (summarized in 2C and 2E).
Specimen 2A 2-cell mz-/-: embryonic day 1.0; Eedtm1d(EUCOMM)Wtsi/Eedtm1d(EUCOMM)Wtsi (more )
Note: Tg(Zp3-cre)93Knw and Tg(Prm-cre)58Og were used to delete the Eed floxed allele in both the female and male germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS2: 2-cell stage embryo Absent 2A mz-/- Devoid of both Eed and H3K27me3 (summarized in 2C and 2E).
Specimen 2-S1B 4-cell fl/fl: embryonic day 2.0; Eedtm1c(EUCOMM)Wtsi/Eedtm1c(EUCOMM)Wtsi (more )
Note: Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS3: 4-cell stage embryo Present 2-S1B 4-cell fl/fl
Specimen 2-S1B 4-cell fl/- or -/-: embryonic day 2.0; Eedtm1d(EUCOMM)Wtsi/? (more )
Note: Tg(Prm-cre)58Og was used to delete the Eed floxed allele in the male germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS3: 4-cell stage embryo Present 2-S1B 4-cell fl/- or -/- A subset of embryos displayed reduced Eed and H3K27me3 levels (summarized in 2C and 2E).
Specimen 2-S1B 4-cell m-/-: embryonic day 2.0; Eedtm1d(EUCOMM)Wtsi/Eed+ (more )
Note: Tg(Zp3-cre)93Knw was used to delete the Eed floxed allele in the female germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS3: 4-cell stage embryo Trace 2-S1B 4-cell m-/- Zygotic Eed transcription initiated (summarized in 2C and 2E).
Specimen 2-S1B 4-cell mz-/-: embryonic day 2.0; Eedtm1d(EUCOMM)Wtsi/Eedtm1d(EUCOMM)Wtsi (more )
Note: Tg(Zp3-cre)93Knw and Tg(Prm-cre)58Og were used to delete the Eed floxed allele in both the female and male germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS3: 4-cell stage embryo Absent 2-S1B 4-cell mz-/-
Specimen 2-S1B 8-cell fl/fl: embryonic day 2.5; Eedtm1c(EUCOMM)Wtsi/Eedtm1c(EUCOMM)Wtsi (more )
Note: Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS3: 8-cell stage embryo Present 2-S1B 8-cell fl/fl
Specimen 2-S1B 8-cell fl/- or -/-: embryonic day 2.5; Eedtm1d(EUCOMM)Wtsi/? (more )
Note: Tg(Prm-cre)58Og was used to delete the Eed floxed allele in the male germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS3: 8-cell stage embryo Present 2-S1B 8-cell fl/- or -/- Highly variable Eed and H3K27me3 levels, suggesting further differentiation of the two genotypes (summarized in 2C and 2E).
Specimen 2-S1B 8-cell m-/-: embryonic day 2.5; Eedtm1d(EUCOMM)Wtsi/Eed+ (more )
Note: Tg(Zp3-cre)93Knw was used to delete the Eed floxed allele in the female germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS3: 8-cell stage embryo Weak 2-S1B 8-cell m-/- In agreement with increasing zygotic Eed expression, Eed m-/- embryos displayed higher levels of Eed and H3K27me3 than the corresponding Eed mz-/- embryos (summarized in 2C and 2E).
Specimen 2-S1B 8-cell mz-/-: embryonic day 2.5; Eedtm1d(EUCOMM)Wtsi/Eedtm1d(EUCOMM)Wtsi (more )
Note: Tg(Zp3-cre)93Knw and Tg(Prm-cre)58Og were used to delete the Eed floxed allele in both the female and male germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS3: 8-cell stage embryo Absent 2-S1B 8-cell mz-/-
Specimen 2B 16-cell fl/fl: embryonic day 3.0; Eedtm1c(EUCOMM)Wtsi/Eedtm1c(EUCOMM)Wtsi (more )
Note: Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS4: compacted morula Present 2B fl/fl
Specimen 2B 16-cell fl/-: embryonic day 3.0; Eedtm1c(EUCOMM)Wtsi/Eedtm1d(EUCOMM)Wtsi (more )
Note: Tg(Prm-cre)58Og was used to delete the Eed floxed allele in the male germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS4: compacted morula Present 2B fl/- Statistically indistinguishable from Eed fl/fl embryos (summarized in 2C and 2E).
Specimen 2B 16-cell -/-: embryonic day 3.0; Eedtm1d(EUCOMM)Wtsi/Eedtm1d(EUCOMM)Wtsi (more )
Note: Tg(Prm-cre)58Og was used to delete the Eed floxed allele in the male germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS4: compacted morula Present 2B -/- Expressed at a lower levels compared with the Eed fl/fl controls (summarized in 2C and 2E).
Specimen 2B 16-cell m-/-: embryonic day 3.0; Eedtm1d(EUCOMM)Wtsi/Eed+ (more )
Note: Tg(Zp3-cre)93Knw was used to delete the Eed floxed allele in the female germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS4: compacted morula Weak 2B m-/- Continued to display higher levels of Eed and H3K27me3 than the Eed mz-/- embryos, but lower levels than Eed fl/fl embryos (summarized in 2C and 2E).
Specimen 2B 16-cell mz-/-: embryonic day 3.0; Eedtm1d(EUCOMM)Wtsi/Eedtm1d(EUCOMM)Wtsi (more )
Note: Tg(Zp3-cre)93Knw and Tg(Prm-cre)58Og were used to delete the Eed floxed allele in both the female and male germline. Double labeled: red - Eed; green - H3K27me3.
Structure Level Pattern Image Note
TS4: compacted morula Absent 2B mz-/-
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04/23/2024
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