however, the assocatobetweeCux1 and apoptoss remans to be elucdat

however, the assocatobetweeCux1 and apoptoss remans for being elucdated.Expermental Procedures Anmals Pkd1cond mce The Pkd1cond mouse lnehas beedescrbed prevously.Ths mouse lnehas loxstes flankng exons 2 via four from the murne Pkd1 gene, therefore allowng the nactvatoof the gene specfcally the tssue of nterest.The Pkd1cond allele s totally functonal and mcehomozygous for ths allele are vable andhealthy.hoxb7 Cre mce Thehoxb7 Cre transgenc mouse lne 13Amc Jwas bought from JacksoLaboratory as well as stock colones are mantaned on the Unversty of Kansas Medcal Center.The Cre recombnase ths transgenc mouse s expressed beneath the manage of your mousehoxb7 enhancer and promoter.The expressoofhoxb7 Cre cabe detected the mesonephrc duct of your kdney as early as embryonc day 9.5.Thereafter, Cre expressocabe seethe mesonephrc duct dervatves with the kdney, whch nclude the collectng duct and the ureteral epthela.Breedng technique We crossed mceheterozygous for the Pkd1cond allele tohoxb7 Cre mce.
Brother sster matngs had been set ubetweeF1 progenyhavng the Pkd1cond wt,Hoxb7 Cre genotype.A total of 59 out selleck of 266 pups have been Pkd1CD.All protocols were approved by the Unversty of Kansas Medcal Center Anmal Care and Use Commttee.The Unversty of Kansas Medcal Center s completely accredted from the AmercaAssocatoof the Accredtatoof Laboratory Anmal Care.Characterzatoof cystc phenotype We analyzed the offsprng of F1 crosses at postnatal day 0, P7, and P15 phases.Kdneys wereharvested from newborn, 7 day previous, 12 day previous and 15 day old mce.Kdneys were weghed plus the complete kdney weght was measured being a percentage of entire body weght for every mouse.harvested kdneys had been fxed 4% paraformaldehyde and embedded paraffn.Sectons of cystc kdneys, staned wthhaematoxyland eosn, had been utzed to stage the cysts, as descrbed prevously.Cysts wth uto 50 cyst lnng epthelal cells had been defned as early stage, ntermedate stage f there were 51 200 cyst lnng epthelal cells, and superior stage cysts f there were extra tha200 cyst lnng epthelal cells.
The cystc ndex was measured H E staned cystc kdney sectons usng mageJ program.Serum chemstry Blood was collected by exsangunaton, followng decaptatoand mmedately centrfuged at 2000 g to acquire serum.Blood urea ntrogethese samples was measured usng QuantChrom Urea Assay Kt.mmunofluorescence Tubastatin A mmunofluorescence was carried out as prevously descrbed.Kdney sectons were ncubated wth 1M ammonum chlorde to quench autofluorescence, washed

Leave a Reply

Your email address will not be published. Required fields are marked *


You may use these HTML tags and attributes: <a href="" title=""> <abbr title=""> <acronym title=""> <b> <blockquote cite=""> <cite> <code> <del datetime=""> <em> <i> <q cite=""> <strike> <strong>