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Antibody-mediated depletion of intestinal myofibroblasts


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#1 Myofibroblast

Myofibroblast

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Posted 30 March 2014 - 01:37 PM

Hello everyone, 

 

I need a little help with this, as I am new to this technique. I would like to use iSEMF specific antibodies for their in vivo depletion. However, several problems arise: 

 

1) There are no specific markers for iSEMF. Usually to define these cells, IF or WB analysis is performed staining for aSMA, Thy1, vimentin and desmin. 

- what will happen when I'd use anti-aSMA antibodies to deplete iSEMF? Would it end up in targeting all aSMA positive sites, such as SMC or pericytes too?

 

2) iSEMF are surrounded by aboundant ECM in the SI villi and LI crypts. Is it likely that ECM would interfere with tagging of the iSEMF in vivo? 

 

Would this be a good approach: 

Inducible knockout (KO) of genes coding for proteins essential in iSEMF development, such as αSMA, MyoD, CLIC4 and ALK5. Crossing strain Myh11-CreERT2, which expresses inducible Cre-recombinase under the control of mouse smooth muscle myosin heavy chain, with gene-of-interest specific conditional-null mice, would allow for SMC- and iSEMF-specific gene deletion forcing the iSEMF to loose their phenotype?

 

Thank you for a feedback!






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