*2.9. Hair Follicle Protein Extract Preparation for Western Immunoblotting*

HF proteins were extracted from a pool of five HFs and prepared in a radioimmunoprecipitation assay buffer (RIPA) buffer supplemented with 1% protease inhibitor cocktail added (Sigma-Aldrich) using the FastPrep 24TM (MP Biomedicals) six times at 6.0 m/s for 40 s. The obtained lysates were incubated 20 min on ice with vortex-mixing every 5 min. Cell debris were precipitated by centrifugation at 10,000 g for 10 min at 4 ◦C. Proteins in the supernatant were collected and assayed using the Bradford (Bio-Rad, Marne-la-Coquette, France) technique [34].
