Hematoxylin and Eosin Staining of Skin Lesions

Hematoxylin and eosin (H&E) staining was performed as previously described [41 (link)]. Briefly, dorsal skin lesions were obtained from the control (PBS), minoxidil (5%), and EF-2001 (0, 5, and 50 mg/mL) groups, fixed in 10% formalin, and stabilized in 10, 15, and 20% sucrose solution. To embed the fixed skin in a cryomold, the FSC 22 Clear frozen section compound (Cat. no. 3801480; Leica Biosystems, Wetzlar, Germany) was used. Afterward, the cryo-tissues were sectioned, stained using H&E, visualized under an Olympus DP80 microscope, and analyzed using imaging software (Cell Sens 1.8; Olympus Europa SE & Co., KG, Hamburg, Germany).