Quantifying Filaggrin and Transglutaminase 3 Expression
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Corresponding Organization :
Other organizations : L'Oréal (France), National and Kapodistrian University of Athens, Physiogenex (France), University of Ioannina
Variable analysis
- Sterile water
- MDixon + SQ 2%
- M. restricta 24 h supernatant B
- SQOOH at 193 ng/ml in mDixon + SQ 2%
- MDA at 60 ng/ml in mDixon + SQ 2%
- SQOOH at 193 ng/ml + MDA at 60 ng/ml in mDixon + SQ 2%
- Level of expression of the proteins filaggrin and transglutaminase 3
- RHE were treated as previously described for morphology and permeability experiments
- Samples were lysed using a specific buffer with Precellys® lysing kits (Bertin Technologies) designed for soft tissue homogenizing and dosed by bicinchoninic acid (BCA) assay
- Diluted protein lysate was mixed with fluorescent master mix and heated at 37°C for 10 min
- Proteins of interest were immunodetected with primary antibodies targeting FLG or TGM3 and peroxidase-conjugated secondary antibodies (Goat anti-Mouse or Rabbit IgG-HRP)
- Signals were normalized to cofilin immunodetection
- None specified
- Sterile water
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