rabbit polyclonal antibodies against h3  (Proteintech)

Journal: Scientific Reports

Article Title: Gradual chromosomal lagging drive programmed genome elimination in hemiclonal fishes from the genus Hypseleotris

doi: 10.1038/s41598-024-78278-6

Figure Lengend Snippet: Analysis of epigenetic histone modifications during mitosis and micronuclei formation. Whole-mount immunofluorescent staining of gonads from sexual species ( a , b ) and hybrids (all rest) with antibodies against various epigenetic modification stained in red: H3S10P ( a , c , d ), H3S28P ( b , e ), H3K9me3 ( f , g ) and H3T11P ( h ). Accumulation of all analyzed chromatin modifications indicate normal chromosome condensation during prophase and metaphase. In hybrids, misaligned chromosomes (indicated by white arrows) have similar distribution of the H3S10P ( c ), H3S28P ( e ), H3K9me3 ( f ) and H3T11P ( h ) chromatin modifications as the rest of the chromosomes (showed by white arrowheads) suggesting that misaligned chromosomes condense properly during prophase and metaphase. Micronuclei present during metaphases (indicated by thin white arrows) do not accumulate epigenetic histone modification such as H3S10P ( c ), H3K9me3 ( f ). ( d ) Accumulation of H3S10P epigenetic modification in some micronuclei (indicated by thin red arrows) suggest their formation by chromosomal lagging. Some micronuclei do not show accumulation of H3S10P epigenetic modification (indicated by thin white arrows). ( g ) In contrast to the chromatin in the interphase nucleus of the gonial cell, chromatin in micronuclei accumulated H3K9me3 epigenetic modifications indicating its inactivation and heterochromatinization (micronuclei indicated with thin red arrows). Tubulin (stained in green) visualize cytoskeleton components of the spindle ( a , f ). Chromatin is visualized with DAPI in blue. All pictures represent a single gonadal section of 0.4 μm in thickness after 3D immunofluorescent staining. Scale bars = 10 μm.

Article Snippet: We used the following primary antibodies: rabbit polyclonal antibody against DDX4 (concentration 1:50; C1C3, GeneTex) to detect Vasa protein; mouse monoclonal antibodies against alpha tubulin (concentration 1:100; ab7291, Abcam); rabbit polyclonal antibody against H3S10P (concentration 1:100; GTX128116, GeneTex); rabbit polyclonal antibody against H3S28P (concentration 1:100; #9713, Cell Signaling); rabbit polyclonal antibody against H3T11P (concentration 1:100; #9764, Cell Signaling); rabbit polyclonal antibody against H3K9me3 (concentration 1:100; ab8898, Abcam).

Techniques: Staining, Modification

Journal: Scientific Reports

Article Title: Gradual chromosomal lagging drive programmed genome elimination in hemiclonal fishes from the genus Hypseleotris

doi: 10.1038/s41598-024-78278-6

Figure Lengend Snippet: Analysis of epigenetic histone modifications during mitosis and micronuclei formation. Whole-mount immunofluorescent staining of gonads from sexual species ( a , b ) and hybrids (all rest) with antibodies against various epigenetic modification stained in red: H3S10P ( a , c , d ), H3S28P ( b , e ), H3K9me3 ( f , g ) and H3T11P ( h ). Accumulation of all analyzed chromatin modifications indicate normal chromosome condensation during prophase and metaphase. In hybrids, misaligned chromosomes (indicated by white arrows) have similar distribution of the H3S10P ( c ), H3S28P ( e ), H3K9me3 ( f ) and H3T11P ( h ) chromatin modifications as the rest of the chromosomes (showed by white arrowheads) suggesting that misaligned chromosomes condense properly during prophase and metaphase. Micronuclei present during metaphases (indicated by thin white arrows) do not accumulate epigenetic histone modification such as H3S10P ( c ), H3K9me3 ( f ). ( d ) Accumulation of H3S10P epigenetic modification in some micronuclei (indicated by thin red arrows) suggest their formation by chromosomal lagging. Some micronuclei do not show accumulation of H3S10P epigenetic modification (indicated by thin white arrows). ( g ) In contrast to the chromatin in the interphase nucleus of the gonial cell, chromatin in micronuclei accumulated H3K9me3 epigenetic modifications indicating its inactivation and heterochromatinization (micronuclei indicated with thin red arrows). Tubulin (stained in green) visualize cytoskeleton components of the spindle ( a , f ). Chromatin is visualized with DAPI in blue. All pictures represent a single gonadal section of 0.4 μm in thickness after 3D immunofluorescent staining. Scale bars = 10 μm.

Article Snippet: We used the following primary antibodies: rabbit polyclonal antibody against DDX4 (concentration 1:50; C1C3, GeneTex) to detect Vasa protein; mouse monoclonal antibodies against alpha tubulin (concentration 1:100; ab7291, Abcam); rabbit polyclonal antibody against H3S10P (concentration 1:100; GTX128116, GeneTex); rabbit polyclonal antibody against H3S28P (concentration 1:100; #9713, Cell Signaling); rabbit polyclonal antibody against H3T11P (concentration 1:100; #9764, Cell Signaling); rabbit polyclonal antibody against H3K9me3 (concentration 1:100; ab8898, Abcam).

Techniques: Staining, Modification

Journal: Scientific Reports

Article Title: Gradual chromosomal lagging drive programmed genome elimination in hemiclonal fishes from the genus Hypseleotris

doi: 10.1038/s41598-024-78278-6

Figure Lengend Snippet: Schematic overview of selective genome elimination in carp gudgeons. ( a ) Scheme of the reproduction of hybrid individuals and individuals of sexual species, H. bucephala . During gametogenesis in hybrids, H. bucephala genome (red) is eliminated, while H. gymnocephala genome (green) is transmitted to gametes. After fertilization by co-occurring sexual species H. bucephala , hybrid chromosomal composition is restored. ( b ) Suggested simplified scheme of gradual chromosome elimination via micronucleus formation in hybrids. ( c ) During eliminating mitosis all chromosomes accumulate epigenetic chromatin marks H3S10P, H3S28P and H3K9me3. After lagging and inclusion of individual chromosomes in micronuclei, we detected H3S10P modification in at least some micronuclei. In the micronuclei, chromatin accumulates heterochromatin marks.

Article Snippet: We used the following primary antibodies: rabbit polyclonal antibody against DDX4 (concentration 1:50; C1C3, GeneTex) to detect Vasa protein; mouse monoclonal antibodies against alpha tubulin (concentration 1:100; ab7291, Abcam); rabbit polyclonal antibody against H3S10P (concentration 1:100; GTX128116, GeneTex); rabbit polyclonal antibody against H3S28P (concentration 1:100; #9713, Cell Signaling); rabbit polyclonal antibody against H3T11P (concentration 1:100; #9764, Cell Signaling); rabbit polyclonal antibody against H3K9me3 (concentration 1:100; ab8898, Abcam).

Techniques: Modification