2.4. Western Blotting

Protein extraction and immunoblotting were performed as previously described [10,13]. In brief, cells were lysed in protein lysis buffer (M-PERTM mammalian protein extraction buffer; Thermo Scientific, Rockford, IL, USA) and cellular debris was removed by centrifugation. Proteins were quantified, separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), transferred to a nitrocellulose membrane, and immunoblotted using the indicated antibodies [10,13]. ImageJ (1.41o) was used for protein quantification.