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Western blot analysis Cells were lysed in lysis buffer [50 mM Tris-HCl (pH 7.5), 1% NP-40, 0.5% (v/v) sodium deoxycholate, 150 mM NaCl and 1 Halt protease and phosphatase inhibitor mixture (Pierce)] on ice for 1 h

Leucocyte glutathione measurement Measurement of GSH concentration in leucocyte subsets was performed by The Ohio State University Veterinary Clinical Flow Cytometry Service (Columbus, OH, USA)

Step 2: Treatment Site Preparation The injection site is cleansed using medical-grade antiseptic protocols to maintain a sterile treatment environment

Pycnogenol's potent antioxidant and anti-inflammatory activity reduces inflammation for proper skin elasticity and smoothness