Western Blot Analysis of Apoptosis Markers

Western blot was executed as previously described [16 (link)]. Shortly, RIPA Lysis Buffer was adopted to extract total proteins. Later on, protein samples were loaded onto 10% SDS-PAGE gel and then transferred to a polyvinylidene fluoride (PVDF) membrane. Subsequently, the membrane was incubated with primary antibodies against Bax (ab243140, 1:500), Bcl-2 (ab32124, 1:1000), IGF1R (ab131476, 1:1000) and β-actin (ab6276, 1:3000) were procured from Abcam (Cambridge, MA, USA). In addition, secondary antibody (ab6702, 1:3000) was used for western blot analysis. Finally, enhanced chemiluminescence kit (Solarbio) was utilized to visualize the protein bands.