- PA5-20710 - Invitrogen Antibodies ATOH8 antibody

George JJ, Martin-Diaz L, Ojanen MJT, Gasa R, Pesu M, Viiri K
International journal of molecular sciences 2021 Aug 28;22(17)

Place ES, Smith JC
PloS one 2017;12(2):e0171143

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of A-20 cell lysate using a ATOH8 polyclonal antibody (Product # PA5-20710) at (A) 1 and (B) 2 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of ATOH8 in A-20 cell lysate with ATOH8 Polyclonal Antibody (Product # PA5-20710) at (A) 1 and (B) 2 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry of ATOH8 in mouse brain tissue with ATOH8 Polyclonal Antibody (Product # PA5-20710) at 5 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 1 Atoh8 sa1465/sa1465 mutants are morphologically normal, with correct heart looping. a) Structure of the zebrafish atoh8 locus and Atoh8 protein. The positions of the A > T substitution in the atoh8 sa1465 allele, and of Lysine 100 in Atoh8 protein, are indicated with asterisks. The red arrowheads mark the positions of each Methionine in the Atoh8 protein, and the basic (orange) and HLH (yellow) domains are indicated. The truncated protein Atoh8 K100X is the predicted product of atoh8 sa1465 . Human ATOH8 contains additional proline-rich (blue) and serine-rich (green) domains. b) Atoh8 Western blot on products from in vitro transcription/translation reactions (TNT Quick) using atoh8 WT/WT and atoh8 sa1465/sa1465 as templates. Predicted size of Atoh8 = 29.8 kDa. Note that there is a strong nonspecific band at ~31 kDa. The full length membrane can be viewed in S1 Fig . c) qPCR on atoh8 WT/WT and atoh8 sa1465/sa1465 embryos (hereafter labelled as ' WT ' and ' sa1465 ', respectively). * p = 0.028. d) WT and sa1465 embryos at 5 days postfertilisation, showing normal overall body morphology and swimbladder inflation. e) Confocal z-stacks showing WT and sa1465 embryo heart morphology.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 2 No alteration in response to atoh8 or fog1 knockdown in atoh8 sa1465/sa1465 mutants. a) Atoh8 Western blot on products from in vitro transcription/translation reactions (TNT Quick) seeded with atoh8 morpholino at the indicated concentrations. The full length membrane can be viewed in S2 Fig . b) Percentage of WT and sa1465 embryos displaying pericardial oedema at 72 hpf, following injection with the indicated morpholinos. c) Examples of pericardial oedema in WT and sa1465 embryos injected with atoh8, id3, and fog1 morpholinos. Imaged at 72 hpf.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Atoh8 is expressed in FAE in Peyer's patches and is dependent on Rank-RankL signaling. ( A ) H3K27me3 occupancy at CpG islands spanning the promoter and first exon of the Atoh8 gene in organoids treated with RankL (M cells) or treated with Wnt3a and Chir99021 (Crypt/ISCs). Below, pre-mRNA expression of Atoh8 in organoids treated as above (y-axis: normalized tag count, ENR500 = R-spondin 500 ng/mL, R100 = Rankl 100 ng/mL). ( B ) Section of PP from wild type mice stained with Atoh8 antibody. Arrowheads indicating Atoh8 expression in the nuclei of M cells in FAE. ( C ) RT-qPCR analysis of Atoh8 and Gp2 in the FAE and VE from C57BL/6JRj mice ( n = 3). ( D ) Organoids generated from wild type mice were stimulated with 100 ng of RankL for 4 d. Atoh8 and Gp2 expression was examined by RT-qPCR analysis. ( E ) Rank KO organoids and Scrambled organoids generated by lentiCRISPR v2 were incubated with RankL for 4 days, Atoh8 and GP2 expression was analyzed by RT-qPCR. In panels ( C - E ), unpaired two-tailed Student's t -test was performed for three independent experiments, *** p < 0.005; ** p < 0.01.

PA5-20710