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In HNSCC, HGF is majorly secreted via tumor-associated fibroblasts (TAFs) in the tumor microenvironment [80, 81]

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Essential Equipment and Materials Required Equipment: Sterile glass vials or low-binding microcentrifuge tubes Calibrated micropipettes (10-1000 L range) Sterile filter tips Vortex mixer or gentle rocker Analytical balance (0.1 mg precision) Reconstitution Solvents: Sterile water (molecular biology grade) Bacteriostatic water (contains 0.9% benzyl alcohol as preservative) Phosphate buffered saline (PBS, pH 7.4) Dilute acetic acid solution (0.1% v/v) DMSO (for highly hydrophobic peptides) Optional Materials: Sonicator bath pH meter 0.22 m sterile filters All materials must be sterile and free from endotoxins for cell culture applications

The protocol timeline is a guideline, not a rigid mandate