Journal: eLife

Article Title: H3K9me2 orchestrates inheritance of spatial positioning of peripheral heterochromatin through mitosis

doi: 10.7554/eLife.49278

Figure Lengend Snippet: ( A ) Representative immunofluorescent confocal images of murine C2C12 cells illustrating localization of H3K9me2- and H3K9me2S10p-marked chromatin and Lamin B during different stages of mitosis; DNA visualized with DAPI. Scale bars: 5 μm. ( B ) Magnified images of Interphase and Prophase from panel ( A ) demonstrating detachment of the H3K9me2-chromatin from the nuclear lamina concomitant with H3K9me2S10p phosphorylation; scale bar: 1 μm. ( C ) Representative images of cells progressing through telophase as the layer of peripheral H3K9me2-marked heterochromatin (green) is re-established and nuclear lamina (Lamin B, red) is reassembled; dashed boxes in top panels indicate higher resolution images. Scale bars: 5 μm (top) and 1 μm (bottom panels). ( D ) Magnified images of telophase and daughter cells from panel A demonstrating de-phosphorylated H3K9me2-chromatin (green) assembled at the nuclear lamina (Lamin B, red), while the phosphorylated form (H3K9me2S10p, cyan, enchanced brightness) remains localized in the nuclear interior; scale bar: 1 μm. Dashed lines indicate location of corresponding representative line signal intensity profiles (bottom row).

Article Snippet: Peptide , H3K9me2S10p , EpiCypher , Cat# 12–0093 , IF (1:500).

Techniques: Phospho-proteomics

Journal: eLife

Article Title: H3K9me2 orchestrates inheritance of spatial positioning of peripheral heterochromatin through mitosis

doi: 10.7554/eLife.49278

Figure Lengend Snippet: ( A ) C2C12 cells in prophase stained with H3K9me2S10p antibody (green) preincubated with indicated blocking peptides, counterstained with DAPI (blue). ( B ) Histone peptide array analysis of H3K9me2S10p antibody reactivity with indicated peptides.

Article Snippet: Peptide , H3K9me2S10p , EpiCypher , Cat# 12–0093 , IF (1:500).

Techniques: Staining, Blocking Assay, Peptide Microarray

Journal: eLife

Article Title: H3K9me2 orchestrates inheritance of spatial positioning of peripheral heterochromatin through mitosis

doi: 10.7554/eLife.49278

Figure Lengend Snippet:

Article Snippet: Peptide , H3K9me2S10p , EpiCypher , Cat# 12–0093 , IF (1:500).

Techniques: Peptide Microarray, Modification, Recombinant, Plasmid Preparation, Sequencing, Mutagenesis, Software

Journal: eLife

Article Title: H3K9me2 orchestrates inheritance of spatial positioning of peripheral heterochromatin through mitosis

doi: 10.7554/eLife.49278

Figure Lengend Snippet:

Article Snippet: Peptide , H3S10p , EpiCypher , Cat# 12–0041 , IF (1:500).

Techniques: Peptide Microarray, Modification, Recombinant, Plasmid Preparation, Sequencing, Mutagenesis, Software

Journal: eLife

Article Title: H3K9me2 orchestrates inheritance of spatial positioning of peripheral heterochromatin through mitosis

doi: 10.7554/eLife.49278

Figure Lengend Snippet:

Article Snippet: Peptide , H3K9me0 , EpiCypher , Cat# 12–0001 , IF (1:500).

Techniques: Peptide Microarray, Modification, Recombinant, Plasmid Preparation, Sequencing, Mutagenesis, Software

Journal: Poultry Science

Article Title: Expression of relaxin-3 and its receptors in the hypothalamic-pituitary-ovary axis in layers and broiler breeders

doi: 10.1016/j.psj.2025.105048

Figure Lengend Snippet: Primary and secondary antibodies used for immunofluorescence.

Article Snippet: Antigen retrieval was performed using the heat-induced epitope retrieval method in sodium citrate buffer (pH 6.0, Abcam, Cambridge, Cambridgeshire, United Kingdom) for 15 min. After the slides cooled down, they were washed twice with PBS-Tween 20, pH 7.4 (PBS-T), rinsed with PBS, and blocked with 5 % normal serum in PBS for 1 h. Immunofluorescence staining was performed using RLN3 (Cat No. 26075-1-AP, 1:50) and RXFP1 (Cat No. 18419-1-AP, 1:100) antibodies purchased from Proteintech Group, Inc (Rosemont, IL, USA) and a custom chicken RXFP3 (1:500) antibody (Thermo Fisher Scientific, Rockford, IL).

Techniques: Immunofluorescence

Journal: Poultry Science

Article Title: Expression of relaxin-3 and its receptors in the hypothalamic-pituitary-ovary axis in layers and broiler breeders

doi: 10.1016/j.psj.2025.105048

Figure Lengend Snippet: Gallus gallus -specific primer sequences.

Article Snippet: Antigen retrieval was performed using the heat-induced epitope retrieval method in sodium citrate buffer (pH 6.0, Abcam, Cambridge, Cambridgeshire, United Kingdom) for 15 min. After the slides cooled down, they were washed twice with PBS-Tween 20, pH 7.4 (PBS-T), rinsed with PBS, and blocked with 5 % normal serum in PBS for 1 h. Immunofluorescence staining was performed using RLN3 (Cat No. 26075-1-AP, 1:50) and RXFP1 (Cat No. 18419-1-AP, 1:100) antibodies purchased from Proteintech Group, Inc (Rosemont, IL, USA) and a custom chicken RXFP3 (1:500) antibody (Thermo Fisher Scientific, Rockford, IL).

Techniques: Sequencing

Journal: Poultry Science

Article Title: Expression of relaxin-3 and its receptors in the hypothalamic-pituitary-ovary axis in layers and broiler breeders

doi: 10.1016/j.psj.2025.105048

Figure Lengend Snippet: Sequence of the gBlock used to determine primer efficiencies.

Article Snippet: Antigen retrieval was performed using the heat-induced epitope retrieval method in sodium citrate buffer (pH 6.0, Abcam, Cambridge, Cambridgeshire, United Kingdom) for 15 min. After the slides cooled down, they were washed twice with PBS-Tween 20, pH 7.4 (PBS-T), rinsed with PBS, and blocked with 5 % normal serum in PBS for 1 h. Immunofluorescence staining was performed using RLN3 (Cat No. 26075-1-AP, 1:50) and RXFP1 (Cat No. 18419-1-AP, 1:100) antibodies purchased from Proteintech Group, Inc (Rosemont, IL, USA) and a custom chicken RXFP3 (1:500) antibody (Thermo Fisher Scientific, Rockford, IL).

Techniques: Sequencing

Journal: Poultry Science

Article Title: Expression of relaxin-3 and its receptors in the hypothalamic-pituitary-ovary axis in layers and broiler breeders

doi: 10.1016/j.psj.2025.105048

Figure Lengend Snippet: Antibody validation for immunofluorescence staining. Representative images demonstrate successful immunofluorescence staining using the validated relaxin 3 (RLN3), relaxin family peptide receptor 1 (RXFP1), and relaxin family peptide receptor 3 (RXFP3). Left panel: negative control with the omission of the primary antibodies, showing minimal background fluorescence. Middle panel: isotype control using normal rabbit IgG replacing the primary antibodies. Right panel: Positive staining in 3-5 mm follicles of broiler breeders, confirming antibody specificity. Results are consistent across all follicle sizes. Scale bars = 100 µm. Abbreviations: GC - Granulosa Cells, OC - Oocyte, TE - Theca Externa, TI - Theca Interna.

Article Snippet: Antigen retrieval was performed using the heat-induced epitope retrieval method in sodium citrate buffer (pH 6.0, Abcam, Cambridge, Cambridgeshire, United Kingdom) for 15 min. After the slides cooled down, they were washed twice with PBS-Tween 20, pH 7.4 (PBS-T), rinsed with PBS, and blocked with 5 % normal serum in PBS for 1 h. Immunofluorescence staining was performed using RLN3 (Cat No. 26075-1-AP, 1:50) and RXFP1 (Cat No. 18419-1-AP, 1:100) antibodies purchased from Proteintech Group, Inc (Rosemont, IL, USA) and a custom chicken RXFP3 (1:500) antibody (Thermo Fisher Scientific, Rockford, IL).

Techniques: Biomarker Discovery, Immunofluorescence, Staining, Negative Control, Fluorescence, Control

Journal: Poultry Science

Article Title: Expression of relaxin-3 and its receptors in the hypothalamic-pituitary-ovary axis in layers and broiler breeders

doi: 10.1016/j.psj.2025.105048

Figure Lengend Snippet: Immunofluorescence localization of relaxin (RLN3) in chicken ovarian follicles of layers and broiler breeders. (A) 9-12 mm follicles. Scale bars = 20 µm. (B) 6-8 mm follicles. Scale bars = 100 µm. (C) 3-5 mm follicles. Scale bars = 20 µm. (D) Ovarian Stroma. Scale bars = 50 µm. Abbreviations: GC - Granulosa Cells, OC - Oocyte, PF – Primary Follicle, PMF – Primordial Follicle, TE - Theca Externa, TI - Theca Interna.

Article Snippet: Antigen retrieval was performed using the heat-induced epitope retrieval method in sodium citrate buffer (pH 6.0, Abcam, Cambridge, Cambridgeshire, United Kingdom) for 15 min. After the slides cooled down, they were washed twice with PBS-Tween 20, pH 7.4 (PBS-T), rinsed with PBS, and blocked with 5 % normal serum in PBS for 1 h. Immunofluorescence staining was performed using RLN3 (Cat No. 26075-1-AP, 1:50) and RXFP1 (Cat No. 18419-1-AP, 1:100) antibodies purchased from Proteintech Group, Inc (Rosemont, IL, USA) and a custom chicken RXFP3 (1:500) antibody (Thermo Fisher Scientific, Rockford, IL).

Techniques: Immunofluorescence

Journal: Poultry Science

Article Title: Expression of relaxin-3 and its receptors in the hypothalamic-pituitary-ovary axis in layers and broiler breeders

doi: 10.1016/j.psj.2025.105048

Figure Lengend Snippet: Comparative mRNA expression of relaxin (RLN3), relaxin family peptide 1 (RXFP1), and relaxin family peptide 3 (RXFP3) mRNA between layers and broiler breeders in ovarian cortex and follicles. (A) RLN3 mRNA expression in granulosa cells (GC) along with the cortex (Left) and theca cells (Right) of the 9-12 mm, 6-8 mm, and 3-5 mm follicles. (B) RXFP1 mRNA expression in granulosa along with the cortex (Left) and theca cells (Right). (C) RXFP3 mRNA expression in granulosa cells along with the cortex (Right) and theca cells (Left). The data are presented as fold changes relative to pooled cDNA. Values are expressed as mean ± SEM analyzed by two-way ANOVA ( n = 4 / hen type / tissue type). Values with different letters are significantly different ( p ≤ 0.05) (Capital letters for layers and lowercase for broiler breeders). Outliers were removed from 3 to 5 mm GC from broiler breeder # 2, 9-12 mm TH from layer # 3-, and 6-8-mm TH from layer # 3.

Article Snippet: Antigen retrieval was performed using the heat-induced epitope retrieval method in sodium citrate buffer (pH 6.0, Abcam, Cambridge, Cambridgeshire, United Kingdom) for 15 min. After the slides cooled down, they were washed twice with PBS-Tween 20, pH 7.4 (PBS-T), rinsed with PBS, and blocked with 5 % normal serum in PBS for 1 h. Immunofluorescence staining was performed using RLN3 (Cat No. 26075-1-AP, 1:50) and RXFP1 (Cat No. 18419-1-AP, 1:100) antibodies purchased from Proteintech Group, Inc (Rosemont, IL, USA) and a custom chicken RXFP3 (1:500) antibody (Thermo Fisher Scientific, Rockford, IL).

Techniques: Expressing

Journal: Poultry Science

Article Title: Expression of relaxin-3 and its receptors in the hypothalamic-pituitary-ovary axis in layers and broiler breeders

doi: 10.1016/j.psj.2025.105048

Figure Lengend Snippet: Comparative mRNA expression of relaxin-3 (RLN3), relaxin family peptide 1 (RXFP1), and relaxin family peptide 3 (RXFP3) in the pituitary and hypothalamus of layer and broiler breeder. (A) RLN3 mRNA expression in the pituitary and the hypothalamus. (B) RXFP1 mRNA expression in the pituitary and the hypothalamus (C) RXFP3 mRNA expression in the pituitary and the hypothalamus. The data are presented as fold changes relative to pooled cDNA. Values are expressed as mean ± SEM analyzed by two-way ANOVA ( n = 4 / hen type / tissue type). Values with different letters are significantly different ( p ≤ 0.05).

Article Snippet: Antigen retrieval was performed using the heat-induced epitope retrieval method in sodium citrate buffer (pH 6.0, Abcam, Cambridge, Cambridgeshire, United Kingdom) for 15 min. After the slides cooled down, they were washed twice with PBS-Tween 20, pH 7.4 (PBS-T), rinsed with PBS, and blocked with 5 % normal serum in PBS for 1 h. Immunofluorescence staining was performed using RLN3 (Cat No. 26075-1-AP, 1:50) and RXFP1 (Cat No. 18419-1-AP, 1:100) antibodies purchased from Proteintech Group, Inc (Rosemont, IL, USA) and a custom chicken RXFP3 (1:500) antibody (Thermo Fisher Scientific, Rockford, IL).

Techniques: Expressing