Production and Characterization of Monoclonal antibodies to Xenopus proteins

Horr, Brett; Kurtz, Ryan; Pandey, Ankit; Hoffstrom, Benjamin G.; Schock, Elizabeth N.; LaBonne, Carole; Alfandari, Dominique

doi:10.1101/2022.12.06.519341

Cited by 2 publications

(5 citation statements)

References 27 publications

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“…The rat monoclonal to flag tag (Agilent Cat# 200474, RRID:AB_10597743). The monoclonal to X. laevis Sox3 (DSHB Cat# DA5H6, RRID:AB_2876376) was developed in the laboratory (Horr et al, 2023) against the full length X. laevis protein produced in Hek293T cells. It has been characterized by Western blot and immunofluorescence (R24OD21485).…”

Section: Antibodiesmentioning

confidence: 99%

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ADAM11 a novel regulator of Wnt and BMP4 signaling in neural crest and cancer

Pandey,

Cousin,

Horr

et al. 2023

Front. Cell Dev. Biol.

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Introduction: Cranial neural crest (CNC) cells are induced at the border of the neural plate by a combination of FGF, Wnt, and BMP4 signaling. CNC then migrate ventrally and invade ventral structures where they contribute to craniofacial development.Methods: We used loss and gain of function experiments to determine phenotypes associated with the perturbation of Adam11 expression in Xenopus Laevis. Mass spectrometry to identify partners of Adam11 and changes in protein expression in CNC lacking Adam11. We used mouse B16 melanoma to test the function of Adam11 in cancer cells, and published database analysis to study the expression of ADAM11 in human tumors.Results: Here we show that a non-proteolytic ADAM, Adam11, originally identified as a putative tumor suppressor binds to proteins of the Wnt and BMP4 signaling pathway. Mechanistic studies concerning these non-proteolytic ADAM lack almost entirely. We show that Adam11 positively regulates BMP4 signaling while negatively regulating β-catenin activity. In vivo, we show that Adam11 influences the timing of neural tube closure and the proliferation and migration of CNC. Using both human tumor data and mouse B16 melanoma cells, we further show that ADAM11 levels similarly correlate with Wnt or BMP4 activation levels.Discussion: We propose that ADAM11 preserves naïve cells by maintaining low Sox3 and Snail/Slug levels through stimulation of BMP4 and repression of Wnt signaling, while loss of ADAM11 results in increased Wnt signaling, increased proliferation and early epithelium to mesenchyme transition.

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Section: Antibodiesmentioning

confidence: 99%

“…Rat monoclonal against N-cadherin (DSHB Cat# MNCD2, RRID:AB_528119). Mouse monoclonal to X. laevis Adam11.L (3C5) was developed in the lab by injecting a bacterial fusion protein containing the DC (disintegrin and cystein rich) domain and isolating the clone by the methods described in (Horr et al, 2023).…”

Section: Antibodiesmentioning

confidence: 99%

ADAM11 a novel regulator of Wnt and BMP4 signaling in neural crest and cancer

Pandey,

Cousin,

Horr

et al. 2023

Front. Cell Dev. Biol.

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“…The copyright holder for this preprint this version posted June 15, 2023. ; https://doi.org/10. 1101/2023 This work was supported by grant from the NIH to DA (R24-OD21485), and DA and HC (R01DE016289). We thank members of the Alfandari lab for helpful comments, discussion and corrections.…”

Section: Acknowledgmentmentioning

confidence: 99%

“…The rat monoclonal to flag tag (Agilent Cat# 200474, RRID:AB_10597743). The monoclonal to Xenopus laevis Sox3 (DSHB Cat# DA5H6, RRID:AB_2876376) was developed in the laboratory (Horr et al, 2023) against the full length Xenopus laevis protein produced in Hek293T cells. It has been characterized by Western blot and immunofluorescence (R24OD21485).…”

Section: Antibodiesmentioning

confidence: 99%

“…Rat monoclonal against N-cadherin (DSHB Cat# MNCD2, RRID:AB_528119). Mouse monoclonal to Xenopus laevis Adam11.L (3C5) was developed in the lab by injecting a bacterial fusion protein containing the DC (disintegrin and cystein rich) domain and isolating the clone by the methods described in (Horr et al, 2023). The copyright holder for this preprint this version posted June 15, 2023. ; https://doi.org/10.1101/2023.06.13.544797 doi: bioRxiv preprint CCAATGGAGGCATAGTGGCACTG).…”

Section: Antibodiesmentioning

confidence: 99%

See 1 more Smart Citation

ADAM11 a novel regulator of Wnt and BMP4 signaling in neural crest and cancer

Pandey

Cousin

Horr

et al. 2023

Preprint

View full text Add to dashboard Cite

Cranial neural crest (CNC) cells are induced at the border of the neural plate by a combination of FGF, Wnt, and BMP4 signaling. CNC then migrate ventrally and invade ventral structures where they contribute to craniofacial development. Here we show that a non-proteolytic ADAM, Adam11, originally identified as a putative tumor suppressor, binds to proteins of the Wnt and BMP4 signaling pathway. Mechanistic studies concerning these non-proteolytic ADAM lack almost entirely. We show that Adam11 positively regulates BMP4 signaling while negatively regulating beta-catenin activity. By modulating these pathways, Adam11 controls the timing of neural tube closure and the proliferation and migration of CNC. Using both human tumor data and mouse B16 melanoma cells, we further show that ADAM11 levels similarly correlate with Wnt or BMP4 activation levels. We propose that ADAM11 preserves naïve cells by maintaining low Sox3 and Snail/Slug levels through stimulation of BMP4 and repression of Wnt signaling, while loss of ADAM11 results in increased Wnt signaling, increased proliferation, and early epithelium to mesenchyme transition.

show abstract

Production and Characterization of Monoclonal antibodies to Xenopus proteins

Cited by 2 publications

References 27 publications

ADAM11 a novel regulator of Wnt and BMP4 signaling in neural crest and cancer

ADAM11 a novel regulator of Wnt and BMP4 signaling in neural crest and cancer

ADAM11 a novel regulator of Wnt and BMP4 signaling in neural crest and cancer

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