) Evaluation of feasible interferences of Red1-C1 fluorescence with FRET ?Further

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Revisão de 21h27min de 17 de janeiro de 2018 por View0bus (discussão | contribs) () Evaluation of feasible interferences of Red1-C1 fluorescence with FRET ?Further)

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) Evaluation of feasible interferences of Red1-C1 fluorescence with FRET ?Further morphological identification of transfected cells ?Exp #12 control ?a) Co-transfection rate is 100  ; b) Red1-C1 does not interfere with FRET ?Confirmation of Exp #3: caspase 3 is constitutively active in CGCs ?Transfected cells are for essentially the most CGCs?Medium 1 (horse serum) ?4 DIV ?Transfection ?Medium 2 (Neurobasal) ?two DIV ?FRET on fixed OCCsSingle pcDNA-SCAT3 DEVG (FRET manage probe)?Medium 1 (horse serum) ?4 DIV ?Transfection ?Medium 1 (horse serum) ?two DIV ?FRET on fixed OCCs?Medium 1 (horse serum) ?four DIV ?Transfection ?Medium 1 (horse serum) ?2 DIV ?FRET on fixed OCCsSingle pcDNA-SCAT3 DEVG (FRET handle probe)Single pcDNA-SCAT3 DEVD (FRET probe)Ac-DEVD-CMK 100 M?Medium 1 (horse serum) ?four DIV ?Transfection ?Medium two (Neurobasal) ?2 DIV ?FRET on fixed Osure, whereas 6B2 has no effect Total TauFig. 6 Low dose PHF OCCsDouble pcDNA-SCAT3 DEVD (FRET probe) + pHcRed1-CLossi et al. Invest Ophthalmol Vis Sci 1997, 38:72?2.Zhu et changed from 0.43 to 1.11 in HeLa cells transfected with pSCAT3-DEVD and challenged with tumor necrosis factor -cycloheximide, a strong activator of Casp3 [24]. Thus, acceptor photobleaching experiments confi.) Evaluation of possible interferences of Red1-C1 fluorescence with FRET ?Further morphological identification of transfected cells ?Exp #12 control ?a) Co-transfection price is 100  ; b) Red1-C1 doesn't interfere with FRET ?Confirmation of Exp #3: caspase three is constitutively active in CGCs ?Transfected cells are for essentially the most CGCs?Medium 1 (horse serum) ?four DIV ?Transfection ?Medium two (Neurobasal) ?2 DIV ?FRET on fixed OCCsSingle pcDNA-SCAT3 DEVG (FRET handle probe)?Medium 1 (horse serum) ?4 DIV ?Transfection ?Medium 1 (horse serum) ?two DIV ?FRET on fixed OCCs?Medium 1 (horse serum) ?4 DIV ?Transfection ?Medium 1 (horse serum) ?two DIV ?FRET on fixed OCCsSingle pcDNA-SCAT3 DEVG (FRET manage probe)Single pcDNA-SCAT3 DEVD (FRET probe)Ac-DEVD-CMK one hundred M?Medium 1 (horse serum) ?4 DIV ?Transfection ?Medium two (Neurobasal) ?two DIV ?FRET on fixed OCCsDouble pcDNA-SCAT3 DEVD (FRET probe) + pHcRed1-CLossi et al. Molecular Neurodegeneration (2016) 11:Web page six ofTable 1 List of experiments, their rationale and primary outcomes (Continued)12 Double pcDNA-SCAT3 DEVD (FRET probe) + pHcRed1-C1-Surv ?Medium 1 (horse serum) ?4 DIV ransfection ?Medium title= jasp.12117 1 (horse serum) ?2 DIV ?FRET on fixed OCCs None ?Evaluation on the effects of survivin overexpression on basal levels of caspase three activation ?Evaluation of the effect of survivin on cell survival ?Survivin reduces basal caspase three activity ?Survivin promotes cell survival None ?Method control: a) Evaluation of co-transfection efficiency; b) Evaluation of achievable interferences of Red1C1 fluorescence with FRET ?Further morphological identification of transfected cells ?Exp. #14 manage ?a) Co-transfection rate is one hundred  ; b) title= jir.2012.0117 Red1-C1 does not interfere with FRET ?Confirmation of Exp #4: caspase three is constitutively active in CGCs ?Transfected cells are for one of the most CGCs 14 Double pcDNA-SCAT3 DEVD (FRET probe) + pHcRed1-C1-Surv ?Medium 1 (horse serum) ?4 DIV ?Transfection ?Medium 2 (Neurobasal) ?two DIV ?FRET on fixed OCCs ?Medium 1 (horse serum) ?4 DIV ?Transfection ?Medium 2 (Neurobasal) ?1 DIV ?FRET on alive OCCs None ?Evaluation on the effects of survivin overexpression on basal levels of caspase three activation ?Evaluation from the impact of survivin on cell survival ?Survivin reduces caspase three activity ?Survivin promotes cell survival ?25 mM KCl ?1 mM NMDA ?1 mM KA ?one hundred?00 M A23187 ?50 M?five mM H2O2 ?60 mM KCl ?one hundred mM H2O2 ?Measurement of caspase three activity right after induction of apoptosis ?A subpopulation of CGCs is insensitive to apoptosis induction and will not display alterations in ECFPem/Venusem ?Other cells display indicators of sufferance ?Genuine time monitoring of caspase 3 in basal circumstances ?Measurement of caspase three activity right after depolarization or oxidative strain ?Caspase three activity is often measured in true time experiments ?Enhance of ECFPem/Venusem just after K+ depolarization but not cell death ?Only tendency to improve of ECFPem/Venusem soon after oxidative pressure.