Anti-Nkx2.5 antibody GTX128357 from GeneTex

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Developmental biology
February 01 2021

Fig 1: Mef2ca+/−;mef2cb−/−mutant embryos have reduced CM and changes in SHF gene expression. (A,B) Embryos from a cross of mef2ca+/−;mef2cb+/−;Tg(myl7:EGFP) and mef2cb+/−;Tg(myl7:EGFP);Tg(myl7:nucDsRed) were fixed and immunostained with GFP (green) and RFP (red) antibodies at 1 dpf or imaged live at 3 and 5 dpf and CM nuclei counted. Confocal stacks are shown on red channel only at 1 dpf for clarity (A). Nuclei number of mef2ca+/−;mef2cb−/− (red bars) was significantly lower than in siblings (blue bars) at all stages (Unpaired Student’s t-test, P-values indicated on graph). Values are mean ± S.E.M. N shown on bars. (C,D) Embryos from a cross of mef2ca+/−;mef2cb+/−;Tg(myl7:EGFP) to mef2cb+/− at 28 hpf (C) and 3 dpf (D) hybridised for ltbp3 mRNA (Fast Red) and myl7:EGFP detected with anti-GFP antibody. Hearts of mef2ca+/−;mef2cb−/− showed less differentiated myocardium (myl7:EGFP) co-expressing ltbp3 mRNA and ltbp3 levels outside the heart were also reduced (C), but by 3 dpf little difference was seen between genotypes (D). (E,F) In situ mRNA hybridisation for nkx2.5 (blue) and myl7 (red) in embryos from a cross of mef2ca+/−;mef2cb+/− and mef2cb−/− at 28 hpf (E, dorsal view) and 52 hpf (F, ventral view, same embryos shown in top panel for nkx2.5 only and in bottom panel for nkx2.5 and myl7). nkx2.5 mRNA was upregulated slightly at 28 hpf but was clearly upregulated throughout the ventricle and down in the atrium at 52 hpf, beyond the normal strong expression in AVC (white arrowhead) and the arterial pole (yellow arrowhead). Dotted line (bottom panel, F) marks chambers. (G) Confocal stack of immunodetection of Nkx2.5 and GFP (myl7:EGFP) of hearts of mef2ca+/−;mef2cb+/− and sibling at 28 hpf in dorsal view showing Nkx2.5 upregulation in the ventricular region of the heart tube (white arrow) and in the arterial pole (yellow arrowhead). Scale bars = 50 μm.
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Developmental biology
February 01 2021

Fig 2: CM addition, hypertrophy and proliferation are involved in ventricular growth between 3 and 5 dpf. (A) Three Optical sections of 5 dpf Tg(myl7:GAL4FF)hu6531;Tg(5XUAS:RFP)zf83;Tg(UAS:h2az2a-GFP)hu6851 larval heart immunostained for GFP (green), RFP (red) and PH3 (magenta) to reveal the sole PH3+nucGFP+ CM detected in this heart (arrow). Note the additional PH3+nucGFP− cells enclosed by the myocardial layer (arrowheads). (B) Number of PH3+ CMs/ventricle at 3 dpf, 4 dpf and 5 dpf. Mean ± SEM. Each dot represents an animal. (C,D)Tg(myl7:GAL4FF)hu6531;Tg(UAS:h2az2a-GFP)hu6851 embryos were exposed to EdU between 4 and 5 dpf. A short confocal stack reveals EdU-labelled CMs (arrowheads, C), which were quantified as the fraction of EdU+ nuclei in H2A-GFP-labelled CMs in ventricle, atrium and whole heart (D). (E,F) Confocal images of anterior ventricle and OFT region of a Tg(nkx2.5:galFF);Tg(UAS:h2a-gfp);Tg(myl7:nucDsRed) larva imaged at 3 and 5 dpf showing a nucGFP+;nucDsRed− cell population (white bracket) adjacent to the OFT region (arrow) at each stage (E). nucGFP+ cells were counted in the bracketed areas (F). (G,H) Confocal stacks of embryos of 3.5 (G) and 6 dpf (H) carrying Tg(myl7:EGFP)twu26 and Tg(–5.1myl7:DsRed2-NLS)f2. G′ and H′ show in greater detail the boxed OFT region. EGFP+ dsRed− cells are clearly present at 3.5 dpf indicating recently differentiated CMs (arrowheads), whereas by 6 dpf all cells seem to be positive for both EGFP and DsRed. (I) Schematic diagram of the Kaede conversion experiment. (J) Confocal stacks of hearts 5 dpf embryos carrying Tg(myl7:GAL4FF)hu6531and Tg(UAS:Kaede)rk8 in which Kaede was converted from green to red at 3 dpf. Transmitted light of the scanned area is shown in J’. Cells expressing green Kaede but no or little red Kaede are marked by white arrow (OFT and ventricle nearby), light blue arrow (AVC), and yellow arrow (ventricle apex). Note, atrial cell expressing green Kaede only (asterisk) was found only in this embryo. Scale bars = 50 μm, except A and E = 20 μm.

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Anti-Nkx2.5 antibody

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GeneTex's Anti-Nkx2.5 antibody is a Rabbit Polyclonal antibody. The Anti-Nkx2.5 antibody was generated using NK2 homeobox 5, nk2.5, nkx2-5, sb, eu581, zgc, 111912 as the antigen. The Anti-Nkx2.5 antibody from GeneTex is a Rabbit Polyclonal antibody to zebrafish Nkx2.5. The Anti-Nkx2.5 antibody Recognizes Zebrafish. The Anti-Nkx2.5 antibody has been validated for the following Applications: IHC-Wm

Developmental biology

Developmental biology

Anti-Nkx2.5 antibody from GeneTex

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GeneTex

Citations (4)

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Anti-Nkx2.5 antibody from GeneTex

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