Aimp2

IHC staining of tumors was performed as described previously [13 (link)]. Primary antibodies were HK2 (Proteintech #22029-1-AP, China), HK2 (Proteintech #66974-1-Ig, China), AIMP2 (Proteintech #10424-1-AP, China), Ki-67 (Cell Signaling Technology #9449, America), Cyto c (Proteintech #10993-1-AP, America), PCNA (Abclonal #a0264, China), LC3 II (Proteintech #18725-1-AP, China), CD8 (Abcam #ab217344), CD163 (Proteintech #16646-1-AP, China), CD206 (Cell Signaling Technology #24595, American). Staining intensity was independently evaluated by two senior pathologists. The immunohistochemical score was calculated based on the positive reaction area and the intensity of staining. Each group quantified at least five different views.

Total proteins were extracted by RIPA buffer (Beyotime, China), lysed from cells or tissues with the addition of protease inhibitors (1:100, CWBIO) and phosphatase inhibitors (1:100, CWBIO), separated by 10% SDS PAGE or 12% SDS PAGE, and then transferred onto PVDF membranes. After incubation with primary antibodies overnight at 4 °C, membranes were washed three times with PBST. These blots were incubated with HRP-conjugated secondary antibody (Proteintech #SA00001-2, America) for 1.5 h at 4 °C. The blots were washed three times with PBST and visualized by enhanced chemiluminescence (Tanon ECL, China). Primary antibodies used for western blot were listed as follows: GAPDH (Proteintech #10494-1-AP, China), HK2 (Proteintech #22029-1-AP, China), AIMP2 (Proteintech #10424-1-AP, China), Cyto c (Proteintech #10993-1-AP, China), Caspase 3/p17/p19 (Proteintech #19677-1-AP, China), Bcl-xL (Abcam #ab32370, America), LC3 (Proteintech #14600-1-AP, China), P62 (Proteintech #18420-1-AP, China), Becline1 (Proteintech #11306-1-AP, China).