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e14 5 fetal liver derived lineage negative cells  (TaKaRa)


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    TaKaRa e14 5 fetal liver derived lineage negative cells
    (A) Schematic representation of MORC3 protein, its different domains and the MommeD41 mutation. (B) Western blot showing MORC3 protein levels in <t>E14.5</t> fetal liver and E18.5 spleen and thymus, two Morc3 +/+ biological replicates. VINCULIN was used as a loading control. (C) Western blot showing MORC3 protein levels in E18.5 thymus, two biological replicates per genotype. VINCULIN was used as a loading control. (D) (top) Representative macroscopic pictures of thymus from E18.5 Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 littermates (scale bars, 0,5cm). (bottom) Total cell count of E18.5 thymus from Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 littermates (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 ). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). (E) Representative flowcytometry plots of T cell subsets in Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 E18.5 thymus. CD4 and CD8 were used as markers to define DN, DP and single positive CD4+ and CD8+. (F) (left) Bar plot depicting the percentage of CD4+CD8+ Double Positive (DP) within CD45.1+ cells (pre-gated on SSC-A/FSC-A, single, live, Lin-) and (right) total cell number in E18.5 thymus (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). (G) (left) Bar plot showing the percentage of CD4-CD8-Double Negative (DN) within CD45.1+ cells (pre-gated on live, Lin-, CD3-) and (right) total cell number in E18.5 thymus (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). For panels D, F and G statistical significance was determined using two-sided t test. (H) Representative flowcytometry plots of DN cells in Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 E18.5 thymus. In the left column, CD25 and CD44 were used as markers to define DN1, DN2, DN3, DN4 T cell subsets within CD4-CD8-CD3-cells. In the middle column, c-kit was used as marker to define ETP and DN1 within CD44+CD25-cells. In the right column, c-kit was used as marker to define DN2a and DN2b within DN2 cells. (I) (top) Bar plot showing the percentage of DN cells in CD45.1+ cells (pre-gated on SSC-A/FSC-A, single, live, Lin-) and (bottom) total cell number in E18.5 thymus. (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). Statistical significance was determined using two-way ANOVA for multiple comparisons. For all figures: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001.
    E14 5 Fetal Liver Derived Lineage Negative Cells, supplied by TaKaRa, used in various techniques. Bioz Stars score: 98/100, based on 973 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e14 5 fetal liver derived lineage negative cells/product/TaKaRa
    Average 98 stars, based on 973 article reviews
    e14 5 fetal liver derived lineage negative cells - by Bioz Stars, 2026-02
    98/100 stars

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    1) Product Images from "The epigenetic reader MORC3 is required for T cell development in the thymus"

    Article Title: The epigenetic reader MORC3 is required for T cell development in the thymus

    Journal: bioRxiv

    doi: 10.1101/2025.03.05.641591

    (A) Schematic representation of MORC3 protein, its different domains and the MommeD41 mutation. (B) Western blot showing MORC3 protein levels in E14.5 fetal liver and E18.5 spleen and thymus, two Morc3 +/+ biological replicates. VINCULIN was used as a loading control. (C) Western blot showing MORC3 protein levels in E18.5 thymus, two biological replicates per genotype. VINCULIN was used as a loading control. (D) (top) Representative macroscopic pictures of thymus from E18.5 Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 littermates (scale bars, 0,5cm). (bottom) Total cell count of E18.5 thymus from Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 littermates (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 ). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). (E) Representative flowcytometry plots of T cell subsets in Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 E18.5 thymus. CD4 and CD8 were used as markers to define DN, DP and single positive CD4+ and CD8+. (F) (left) Bar plot depicting the percentage of CD4+CD8+ Double Positive (DP) within CD45.1+ cells (pre-gated on SSC-A/FSC-A, single, live, Lin-) and (right) total cell number in E18.5 thymus (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). (G) (left) Bar plot showing the percentage of CD4-CD8-Double Negative (DN) within CD45.1+ cells (pre-gated on live, Lin-, CD3-) and (right) total cell number in E18.5 thymus (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). For panels D, F and G statistical significance was determined using two-sided t test. (H) Representative flowcytometry plots of DN cells in Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 E18.5 thymus. In the left column, CD25 and CD44 were used as markers to define DN1, DN2, DN3, DN4 T cell subsets within CD4-CD8-CD3-cells. In the middle column, c-kit was used as marker to define ETP and DN1 within CD44+CD25-cells. In the right column, c-kit was used as marker to define DN2a and DN2b within DN2 cells. (I) (top) Bar plot showing the percentage of DN cells in CD45.1+ cells (pre-gated on SSC-A/FSC-A, single, live, Lin-) and (bottom) total cell number in E18.5 thymus. (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). Statistical significance was determined using two-way ANOVA for multiple comparisons. For all figures: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001.
    Figure Legend Snippet: (A) Schematic representation of MORC3 protein, its different domains and the MommeD41 mutation. (B) Western blot showing MORC3 protein levels in E14.5 fetal liver and E18.5 spleen and thymus, two Morc3 +/+ biological replicates. VINCULIN was used as a loading control. (C) Western blot showing MORC3 protein levels in E18.5 thymus, two biological replicates per genotype. VINCULIN was used as a loading control. (D) (top) Representative macroscopic pictures of thymus from E18.5 Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 littermates (scale bars, 0,5cm). (bottom) Total cell count of E18.5 thymus from Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 littermates (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 ). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). (E) Representative flowcytometry plots of T cell subsets in Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 E18.5 thymus. CD4 and CD8 were used as markers to define DN, DP and single positive CD4+ and CD8+. (F) (left) Bar plot depicting the percentage of CD4+CD8+ Double Positive (DP) within CD45.1+ cells (pre-gated on SSC-A/FSC-A, single, live, Lin-) and (right) total cell number in E18.5 thymus (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). (G) (left) Bar plot showing the percentage of CD4-CD8-Double Negative (DN) within CD45.1+ cells (pre-gated on live, Lin-, CD3-) and (right) total cell number in E18.5 thymus (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). For panels D, F and G statistical significance was determined using two-sided t test. (H) Representative flowcytometry plots of DN cells in Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 E18.5 thymus. In the left column, CD25 and CD44 were used as markers to define DN1, DN2, DN3, DN4 T cell subsets within CD4-CD8-CD3-cells. In the middle column, c-kit was used as marker to define ETP and DN1 within CD44+CD25-cells. In the right column, c-kit was used as marker to define DN2a and DN2b within DN2 cells. (I) (top) Bar plot showing the percentage of DN cells in CD45.1+ cells (pre-gated on SSC-A/FSC-A, single, live, Lin-) and (bottom) total cell number in E18.5 thymus. (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). Statistical significance was determined using two-way ANOVA for multiple comparisons. For all figures: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001.

    Techniques Used: Mutagenesis, Western Blot, Control, Cell Counting, Standard Deviation, Marker

    (A) Schematic illustration of the differentiation of T cells from E14.5 fetal liver-derived lineage negative (Lin- : CD3-, CD4-, B220-, Gr1-, NK1.1-, Cd11c-, TER119-) cells co-cultured on OP9-DL1 cells. (B) Representative flowcytometry plots of Morc3 +/+ and Morc3 MD41/MD41 DN1, DN2, DN3 and DN4 cells generated from E14.5 fetal liver-derived lineage negative cells co-cultured on OP9-DL1 cells for 7 days. (C) (top) Percentage of DN1, DN2, DN3 and DN4 cells within Thy1.1+ cells (pre-gated on live, Lin-, CD45.1+) and (bottom) total cell number (n = 5 biological replicates per genotype, 3 independent experiments); data are presented as mean with dots/triangles as individual values; error bar represents SD. Statistical significance was determined using two-sided t test. (D) RT-qPCR analysis of relative Morc3 , Bcl11a , Spi1 and Tcf7 mRNA levels in Morc3 +/+ and Morc3 MD41/MD41 sorted DN1 cells after 7 days of co-culture (normalized to β-actin; n = 4 biological replicates; data are presented as mean with dots/triangles as individual values; error bar represents SD. Statistical significance was determined using two-sided t test. (E) Representative flowcytometry plots of Morc3 +/+ and Morc3 MD41/MD41 myeloid (Gr1+, Cd11b+), NK (NK1.1+, Gr1-,Cd11b-) and B cells (CD19+, B220+, Gr1-,Cd11b-, NK1.1-, Thy1.1-) generated from E14.5 fetal liver-derived lineage negative cells co-cultured on OP9-DL1 cells for 7 days. All cells were pre-gated on single, live, CD45.1+. (F) (top) Percentage of myeloid, NK and B cells within CD45.1+ cells (pre-gated on single, live) and (bottom) total cell number (n = 4 biological replicates/genotype, 2 independent experiments); data are presented as mean with dots/triangle as individual values; error bar represents SD. Statistical significance was determined using two-sided t test. For all figures: *P≤0.05, **P≤0.01, ***P≤0.001, ****P ≤ 0.0001
    Figure Legend Snippet: (A) Schematic illustration of the differentiation of T cells from E14.5 fetal liver-derived lineage negative (Lin- : CD3-, CD4-, B220-, Gr1-, NK1.1-, Cd11c-, TER119-) cells co-cultured on OP9-DL1 cells. (B) Representative flowcytometry plots of Morc3 +/+ and Morc3 MD41/MD41 DN1, DN2, DN3 and DN4 cells generated from E14.5 fetal liver-derived lineage negative cells co-cultured on OP9-DL1 cells for 7 days. (C) (top) Percentage of DN1, DN2, DN3 and DN4 cells within Thy1.1+ cells (pre-gated on live, Lin-, CD45.1+) and (bottom) total cell number (n = 5 biological replicates per genotype, 3 independent experiments); data are presented as mean with dots/triangles as individual values; error bar represents SD. Statistical significance was determined using two-sided t test. (D) RT-qPCR analysis of relative Morc3 , Bcl11a , Spi1 and Tcf7 mRNA levels in Morc3 +/+ and Morc3 MD41/MD41 sorted DN1 cells after 7 days of co-culture (normalized to β-actin; n = 4 biological replicates; data are presented as mean with dots/triangles as individual values; error bar represents SD. Statistical significance was determined using two-sided t test. (E) Representative flowcytometry plots of Morc3 +/+ and Morc3 MD41/MD41 myeloid (Gr1+, Cd11b+), NK (NK1.1+, Gr1-,Cd11b-) and B cells (CD19+, B220+, Gr1-,Cd11b-, NK1.1-, Thy1.1-) generated from E14.5 fetal liver-derived lineage negative cells co-cultured on OP9-DL1 cells for 7 days. All cells were pre-gated on single, live, CD45.1+. (F) (top) Percentage of myeloid, NK and B cells within CD45.1+ cells (pre-gated on single, live) and (bottom) total cell number (n = 4 biological replicates/genotype, 2 independent experiments); data are presented as mean with dots/triangle as individual values; error bar represents SD. Statistical significance was determined using two-sided t test. For all figures: *P≤0.05, **P≤0.01, ***P≤0.001, ****P ≤ 0.0001

    Techniques Used: Derivative Assay, Cell Culture, Generated, Quantitative RT-PCR, Co-Culture Assay



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    e14 5 fetal liver derived lineage negative cells  (TaKaRa)
    98
    TaKaRa e14 5 fetal liver derived lineage negative cells
    (A) Schematic representation of MORC3 protein, its different domains and the MommeD41 mutation. (B) Western blot showing MORC3 protein levels in <t>E14.5</t> fetal liver and E18.5 spleen and thymus, two Morc3 +/+ biological replicates. VINCULIN was used as a loading control. (C) Western blot showing MORC3 protein levels in E18.5 thymus, two biological replicates per genotype. VINCULIN was used as a loading control. (D) (top) Representative macroscopic pictures of thymus from E18.5 Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 littermates (scale bars, 0,5cm). (bottom) Total cell count of E18.5 thymus from Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 littermates (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 ). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). (E) Representative flowcytometry plots of T cell subsets in Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 E18.5 thymus. CD4 and CD8 were used as markers to define DN, DP and single positive CD4+ and CD8+. (F) (left) Bar plot depicting the percentage of CD4+CD8+ Double Positive (DP) within CD45.1+ cells (pre-gated on SSC-A/FSC-A, single, live, Lin-) and (right) total cell number in E18.5 thymus (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). (G) (left) Bar plot showing the percentage of CD4-CD8-Double Negative (DN) within CD45.1+ cells (pre-gated on live, Lin-, CD3-) and (right) total cell number in E18.5 thymus (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). For panels D, F and G statistical significance was determined using two-sided t test. (H) Representative flowcytometry plots of DN cells in Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 E18.5 thymus. In the left column, CD25 and CD44 were used as markers to define DN1, DN2, DN3, DN4 T cell subsets within CD4-CD8-CD3-cells. In the middle column, c-kit was used as marker to define ETP and DN1 within CD44+CD25-cells. In the right column, c-kit was used as marker to define DN2a and DN2b within DN2 cells. (I) (top) Bar plot showing the percentage of DN cells in CD45.1+ cells (pre-gated on SSC-A/FSC-A, single, live, Lin-) and (bottom) total cell number in E18.5 thymus. (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). Statistical significance was determined using two-way ANOVA for multiple comparisons. For all figures: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001.
    E14 5 Fetal Liver Derived Lineage Negative Cells, supplied by TaKaRa, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e14 5 fetal liver derived lineage negative cells/product/TaKaRa
    Average 98 stars, based on 1 article reviews
    e14 5 fetal liver derived lineage negative cells - by Bioz Stars, 2026-02
    98/100 stars
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    (A) Schematic representation of MORC3 protein, its different domains and the MommeD41 mutation. (B) Western blot showing MORC3 protein levels in E14.5 fetal liver and E18.5 spleen and thymus, two Morc3 +/+ biological replicates. VINCULIN was used as a loading control. (C) Western blot showing MORC3 protein levels in E18.5 thymus, two biological replicates per genotype. VINCULIN was used as a loading control. (D) (top) Representative macroscopic pictures of thymus from E18.5 Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 littermates (scale bars, 0,5cm). (bottom) Total cell count of E18.5 thymus from Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 littermates (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 ). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). (E) Representative flowcytometry plots of T cell subsets in Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 E18.5 thymus. CD4 and CD8 were used as markers to define DN, DP and single positive CD4+ and CD8+. (F) (left) Bar plot depicting the percentage of CD4+CD8+ Double Positive (DP) within CD45.1+ cells (pre-gated on SSC-A/FSC-A, single, live, Lin-) and (right) total cell number in E18.5 thymus (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). (G) (left) Bar plot showing the percentage of CD4-CD8-Double Negative (DN) within CD45.1+ cells (pre-gated on live, Lin-, CD3-) and (right) total cell number in E18.5 thymus (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). For panels D, F and G statistical significance was determined using two-sided t test. (H) Representative flowcytometry plots of DN cells in Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 E18.5 thymus. In the left column, CD25 and CD44 were used as markers to define DN1, DN2, DN3, DN4 T cell subsets within CD4-CD8-CD3-cells. In the middle column, c-kit was used as marker to define ETP and DN1 within CD44+CD25-cells. In the right column, c-kit was used as marker to define DN2a and DN2b within DN2 cells. (I) (top) Bar plot showing the percentage of DN cells in CD45.1+ cells (pre-gated on SSC-A/FSC-A, single, live, Lin-) and (bottom) total cell number in E18.5 thymus. (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). Statistical significance was determined using two-way ANOVA for multiple comparisons. For all figures: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001.

    Journal: bioRxiv

    Article Title: The epigenetic reader MORC3 is required for T cell development in the thymus

    doi: 10.1101/2025.03.05.641591

    Figure Lengend Snippet: (A) Schematic representation of MORC3 protein, its different domains and the MommeD41 mutation. (B) Western blot showing MORC3 protein levels in E14.5 fetal liver and E18.5 spleen and thymus, two Morc3 +/+ biological replicates. VINCULIN was used as a loading control. (C) Western blot showing MORC3 protein levels in E18.5 thymus, two biological replicates per genotype. VINCULIN was used as a loading control. (D) (top) Representative macroscopic pictures of thymus from E18.5 Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 littermates (scale bars, 0,5cm). (bottom) Total cell count of E18.5 thymus from Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 littermates (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 ). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). (E) Representative flowcytometry plots of T cell subsets in Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 E18.5 thymus. CD4 and CD8 were used as markers to define DN, DP and single positive CD4+ and CD8+. (F) (left) Bar plot depicting the percentage of CD4+CD8+ Double Positive (DP) within CD45.1+ cells (pre-gated on SSC-A/FSC-A, single, live, Lin-) and (right) total cell number in E18.5 thymus (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). (G) (left) Bar plot showing the percentage of CD4-CD8-Double Negative (DN) within CD45.1+ cells (pre-gated on live, Lin-, CD3-) and (right) total cell number in E18.5 thymus (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). For panels D, F and G statistical significance was determined using two-sided t test. (H) Representative flowcytometry plots of DN cells in Morc3 +/+ , Morc3 +/MD41 and Morc3 MD41/MD41 E18.5 thymus. In the left column, CD25 and CD44 were used as markers to define DN1, DN2, DN3, DN4 T cell subsets within CD4-CD8-CD3-cells. In the middle column, c-kit was used as marker to define ETP and DN1 within CD44+CD25-cells. In the right column, c-kit was used as marker to define DN2a and DN2b within DN2 cells. (I) (top) Bar plot showing the percentage of DN cells in CD45.1+ cells (pre-gated on SSC-A/FSC-A, single, live, Lin-) and (bottom) total cell number in E18.5 thymus. (n = 8 Morc3 +/+ , n = 8 Morc3 +/MD41 , n = 5 Morc3 MD41/MD41 , 3 independent experiments). Data are presented as mean with dot, square or triangle as individual values; error bar represents standard deviation (SD). Statistical significance was determined using two-way ANOVA for multiple comparisons. For all figures: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, **** P ≤ 0.0001.

    Article Snippet: E14.5 fetal liver-derived lineage negative cells were transduced using RetroNectin (T100B, Takara Bio Inc.) coated wells following the manufacturer’s instructions.

    Techniques: Mutagenesis, Western Blot, Control, Cell Counting, Standard Deviation, Marker

    (A) Schematic illustration of the differentiation of T cells from E14.5 fetal liver-derived lineage negative (Lin- : CD3-, CD4-, B220-, Gr1-, NK1.1-, Cd11c-, TER119-) cells co-cultured on OP9-DL1 cells. (B) Representative flowcytometry plots of Morc3 +/+ and Morc3 MD41/MD41 DN1, DN2, DN3 and DN4 cells generated from E14.5 fetal liver-derived lineage negative cells co-cultured on OP9-DL1 cells for 7 days. (C) (top) Percentage of DN1, DN2, DN3 and DN4 cells within Thy1.1+ cells (pre-gated on live, Lin-, CD45.1+) and (bottom) total cell number (n = 5 biological replicates per genotype, 3 independent experiments); data are presented as mean with dots/triangles as individual values; error bar represents SD. Statistical significance was determined using two-sided t test. (D) RT-qPCR analysis of relative Morc3 , Bcl11a , Spi1 and Tcf7 mRNA levels in Morc3 +/+ and Morc3 MD41/MD41 sorted DN1 cells after 7 days of co-culture (normalized to β-actin; n = 4 biological replicates; data are presented as mean with dots/triangles as individual values; error bar represents SD. Statistical significance was determined using two-sided t test. (E) Representative flowcytometry plots of Morc3 +/+ and Morc3 MD41/MD41 myeloid (Gr1+, Cd11b+), NK (NK1.1+, Gr1-,Cd11b-) and B cells (CD19+, B220+, Gr1-,Cd11b-, NK1.1-, Thy1.1-) generated from E14.5 fetal liver-derived lineage negative cells co-cultured on OP9-DL1 cells for 7 days. All cells were pre-gated on single, live, CD45.1+. (F) (top) Percentage of myeloid, NK and B cells within CD45.1+ cells (pre-gated on single, live) and (bottom) total cell number (n = 4 biological replicates/genotype, 2 independent experiments); data are presented as mean with dots/triangle as individual values; error bar represents SD. Statistical significance was determined using two-sided t test. For all figures: *P≤0.05, **P≤0.01, ***P≤0.001, ****P ≤ 0.0001

    Journal: bioRxiv

    Article Title: The epigenetic reader MORC3 is required for T cell development in the thymus

    doi: 10.1101/2025.03.05.641591

    Figure Lengend Snippet: (A) Schematic illustration of the differentiation of T cells from E14.5 fetal liver-derived lineage negative (Lin- : CD3-, CD4-, B220-, Gr1-, NK1.1-, Cd11c-, TER119-) cells co-cultured on OP9-DL1 cells. (B) Representative flowcytometry plots of Morc3 +/+ and Morc3 MD41/MD41 DN1, DN2, DN3 and DN4 cells generated from E14.5 fetal liver-derived lineage negative cells co-cultured on OP9-DL1 cells for 7 days. (C) (top) Percentage of DN1, DN2, DN3 and DN4 cells within Thy1.1+ cells (pre-gated on live, Lin-, CD45.1+) and (bottom) total cell number (n = 5 biological replicates per genotype, 3 independent experiments); data are presented as mean with dots/triangles as individual values; error bar represents SD. Statistical significance was determined using two-sided t test. (D) RT-qPCR analysis of relative Morc3 , Bcl11a , Spi1 and Tcf7 mRNA levels in Morc3 +/+ and Morc3 MD41/MD41 sorted DN1 cells after 7 days of co-culture (normalized to β-actin; n = 4 biological replicates; data are presented as mean with dots/triangles as individual values; error bar represents SD. Statistical significance was determined using two-sided t test. (E) Representative flowcytometry plots of Morc3 +/+ and Morc3 MD41/MD41 myeloid (Gr1+, Cd11b+), NK (NK1.1+, Gr1-,Cd11b-) and B cells (CD19+, B220+, Gr1-,Cd11b-, NK1.1-, Thy1.1-) generated from E14.5 fetal liver-derived lineage negative cells co-cultured on OP9-DL1 cells for 7 days. All cells were pre-gated on single, live, CD45.1+. (F) (top) Percentage of myeloid, NK and B cells within CD45.1+ cells (pre-gated on single, live) and (bottom) total cell number (n = 4 biological replicates/genotype, 2 independent experiments); data are presented as mean with dots/triangle as individual values; error bar represents SD. Statistical significance was determined using two-sided t test. For all figures: *P≤0.05, **P≤0.01, ***P≤0.001, ****P ≤ 0.0001

    Article Snippet: E14.5 fetal liver-derived lineage negative cells were transduced using RetroNectin (T100B, Takara Bio Inc.) coated wells following the manufacturer’s instructions.

    Techniques: Derivative Assay, Cell Culture, Generated, Quantitative RT-PCR, Co-Culture Assay