Western dot kit

Levels of proteins engaged in the apoptotic process (caspase-8, procaspase-9, procaspase-3), were determined by western blot anaysis. A sample was separated with SDS-PAGE and transferred to a PVDF membrane (Amersham, GE Healthcare Life Sciences, Freiburg, Germany) using a vertical electrophoresis apparatus Mini-PROTEAN ^® 3 Cell (BioRad, Hercules, CA, USA.). Proteins were immunoblotted with the primary monoclonal antibodies: anti- aspase-8, anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Thermo Fisher Scientific, Waltham, MA, USA), anticaspase 3 and 9 (Cell Signaling, Leiden, The Netherlands). GAPDH was amplified as an internal control. Protein bands were visualized with the Bio Imaging System (DNR Lumi BIS, Jerusalem, Israel), using the fluorescent method of the Western Dot Kit (Thermo Fisher Scientific). Protein bands were characterized using ImagineR (Sun Microsystems, Santa Clara, CA, USA) analysis software.

The levels of proteins engaged in the apoptotic process (caspase 8, procaspase 9, and procaspase 3) and autophagy, i.e., LC3-II, were tested. A sample was separated with SDS-PAGE and transferred to a PVDF membrane (Amersham, GE Healthcare Life Sciences, Freiburg, Germany) using vertical electrophoresis apparatus Mini-PROTEAN^® 3 Cell (BioRad, Hercules, CA, USA). The proteins were immunoblotted with the primary monoclonal antibodies: anti-caspase 8, anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Thermo Fisher Scientific, Waltham, MA, USA), and anti-caspase 3 and 9 (Cell Signaling, Leiden, The Netherlands). GAPDH was amplified as an internal control. Protein bands were visualized with a Bio Imaging System (DNR Lumi BIS, Jerusalem, Israel) using the fluorescent method of a WesternDot kit (Thermo Fisher Scientific). Protein bands were characterized using the ImagineR (Sun Microsystems, Santa Clara, CA, USA) analysis software [50 (link)].