SDS ges for Western bot nysis. Smpes were processed by eectrophoreseseec trobot s described bovenyzed by Western botting using rbbit ntiMTMMP IgG . gm; Snt Cruz Biotechnoogy. The experiment ws performed times, with simir resuts obtined in ech TAK-875 experiment. Sttistic nysis.dt were obtined from t est iepeent experiments, ech performed in tripicte. Sttistic significnce ws determined using Student’s t test. P vues ess thn . were considered significnt. RESUTS H oigoscchride stimution of DMTS ure . Hyuronn H oigoscchridemedited stimution of ggrecnse mRN expression in chorocytes derived from different yers of rticur crtige.

Chorocytes were isoted from fu thickness rticur crtige, the upper yers upper of rticur crtige, or the middetodeep zone yers owerof rticur crtigecutured forhours in the presence or bsence of gm of H oigoscchrides. The fod chnge in DMTSDMTS B mRN copy number in controtreted cuturesDMTS mRN expression. Primer sets for retime RTPCR mpifictionquntifiction of bovine DMTS, DMTS,GPDH were ws determined. Vues re the men SD of tripicte cutures. designed. mpifiction efficiencies for ech primer set were determined owing the prison of mRN copy numbers between tretedcontro cutures, which were normized to GPDH. Chorocytes were incubted forhours uer serumfree coitions in the bsence of H oigoscchrides or for, or with H oigoscchrides so produced sttisticy significnt increse in DMTS mRN expression by hours, reching mxim .fod increse thours.

H oigoscchridemedited stimution of ggrecnse mRN ws trnsient,copy numbers hd returned to bs eves t the hour time pointhours in the presence of gm of H oigosc uresB.  Etoposide chridesthen processed for tot RN. s ery s hours of incubtion with H oigoscchrides, there ws .fod increse in DMTS mRN copy number s pred to un treted contro chorocytes ure . This effect of H oigoscchrides ws timedepeent, with the mx imum enhncement observed t the hour time point .fod increse. s shown in ure B, tretment H oigoscchrides effected concentrtion depeent increse in ggrecnse, reching sttistic significnce t gm foowinghours of tretmentreching gm foowinghours of tretment ures CD. s contro, the sme H oigo scchrides were predigested with choroitinse BC to generte H discchrides. Incubtion of choro cytes with H discchrides t concentrtion of OIGOSCCHRIDE STIMUTION OF GGRECNSESure Time courseconcentrtion depeency of ggrecnse protein stimution by hyuronn H oigoscchrides. Shown re Western bots of concentrted coitioned medium probed with ntibodies specific for DMTS, detecting DMTS species tkd, kd, kdC or specific for DMTS protein BD .

Chorocytes cutured forhours without tretment contro Ctr or cutured fororhours in the Semagacestat Y-secretase inhibitor presence of gm of H oigoscchrides. CD, Chorocytes cutured forhours without tretment or with gm of H oigoscchrides. Representtive Western bots from of seprte experiments re shown t the top. Densitometric nyses of the respective Western bots re shown t the bottom. For the DMTS dt, soid brs show p, shded brs show p,open brs show p. Vues re the normized men SD fod chnge in pixe intensity for ech psychiatrists coition n seprte experiments. gm resuted in no stimution of DMTS or DMTS mRN ures CD. These resuts suggest tht the ggrecnse stimutory ctivity ob served is due to regent tht is sensitive to choroiti nse digestionnot contminnt. The mximum eves of stimution of DMTSDMTS mRN iuced by H oigoscchrides . respectivey ure were substnti, but were ower thn the vues obtined in pre cutures treted with ngm of I forhours. In these ces, I tretment resuted in fod increse in DMTS.fod increse in DMTS mRN s pred to control Semagacestat 425386-60-3 chorocytes dt not shown. To determine whether prticur subpoputions of chorocytes respo differenty to H oigoscch rides, rticur chorocytes were isoted from the upper yers the ower, middedeep zone yers of bovine crtige.