Mouse monoclonal antibody against β actin

Cell extracts were separated on SDS-polyacrylamide gels, and then the proteins were transferred to a nitrocellulose membrane and incubated with the rabbit polyclonal antibodies: anti-LC3B (1:500; Cell Signaling Technology, Danvers, MA, USA), anti-Beclin1 (1:500; Cell Signaling Technology), anti-ATG5 (1:500; Cell Signaling Technology), anti-caspase3 (1:500; Cell Signaling Technology), anti-cleaved-caspase3 (1:500; Cell Signaling Technology), anti-PARP (1:500; Cell Signaling Technology), anti-cleaved-PARP (1:500; Cell Signaling Technology), anti-JNK (1:500; Cell Signaling Technology), anti-p-JNK (1:500; Cell Signaling Technology), anti-AKT (1:500; Cell Signaling Technology), anti-p-AKT (1:500; Cell Signaling Technology), anti-ERK (1:500; Cell Signaling Technology), anti-p-ERK (1:500; Cell Signaling Technology), anti-p38 (1:500; Cell Signaling Technology), and anti-p-p38 (1:500; Cell Signaling Technology), as well as a mouse monoclonal antibody against β-actin (1:1000; Cell Signaling Technology). Immunoreactive protein bands were detected with Tanon scanning system (Tanon Science & Technology Co., Ltd., Beijing, China).

The rabbit polyclone antibody against RasGRF1 (Cat.No:12958-1-AP) was from Proteintech group (Wuhan, China) for Western blot. The rabbit polyclone antibody against RasGRF1 (Cat.No: GB11856) was from Servicebio company (Wuhan, China) for immunohistochemistry. The mouse monoclonal antibody against β-actin was purchased from Cell signaling Technology, Inc. (Danvers, MA, USA). The labeled HRP secondary antibodies against rabbit and mouse IgG were from Biosharp Life Sciences (Hefei, Anhui, China). Biotin-labeling goat antibody against rabbit IgG and HRP-labeling streptavidin were from Zymed Technologies (San Diego, CA, USA). The diaminobenzidine (DAB) detection kit was from ZSbio.ltd. (Beijing, China). Mimics (miRNA-323-3p, miRNA-375-3p, and miRNA-448-5p), miRNA-323-3p antagomir and negative control were from RiboBio company (Guangzhou, China). RasGRF1 siRNA and scramble siRNA were designed and synthesized by Shanghai Genechem Co., Ltd. (Shanghai, China).