Anti sirpα

Mesoporous silica was purchased from nanoComposix (USA). Shanzhiside methylester was purchased from DESITE (China). DOX and Dialysis Membrane (2 kD) were purchased from Solarbio (China). Cy5, Hoechst 33342, and LysoTracker Green DND-26 were purchased from Yeasen Biotechnology (China). DSPE-PEG-FITC, MW:2000 purchased from Ruixi Biology Co., Ltd (China). Annexin V-FITC apoptosis detection kit was purchased from Beyotime Biotechnology (China). ROS assay kit was purchased from Beyotime Biotechnology (China). JC-1 MitoMP Detection Kit and cell counting kit-8 (CCK-8) were purchased from Dojindo Laboratories (Japan). Penicillin and streptomycin cocktail, fetal bovine serum (FBS), and RPMI-1640 were purchased from Life Technologies (USA). Anti-Bcl-2, anti-Bax, anti-TNF-α and anti-IL-1β, anti-CD47, and anti-SIRPα were manufactured by Cell Signaling Technology (USA). Anti-Ly6c was purchased from Abcam (UK). TNF-α ELISA. IL-1β ELISA was purchased from R&D Systems (China). HRP conjugated goat anti-rabbit IgG and HRP conjugated goat anti-mouse IgG were manufactured by Auragene Biotech (China). TdT in situ apoptotic kit was purchased from R&D Systems (China). Hematoxylin eosin (H&E) and DAPI were purchased from Servicebio Technology (China). Polycarbonate porous membrane syringe filters (200 nm) were provided by Whatman (USA).

Total protein was isolated with RIPA buffer (#P0013, Beyotime, China) and quantified with a BCA kit (#P0009, Beyotime, China). Each sample (50 μg) was separated by 10% SDS-PAGE and transferred to a polyvinylidene difluoride membrane, which was then blocked with 5% BSA and incubated with primary antibodies overnight at 4 °C. Membranes were then rinsed 3 times with PBS containing 0.1% Tween 20 (PBST) and incubated with appropriate secondary antibodies at room temperature for 1 h. Then, membranes were washed with PBST 3 times. Signals were generated with Enhanced Chemiluminescence Substrate (#NEL105001 EA, PerkinElmer) for 1 min before detection with a Bio-Rad ChemiDoc MP System (170-8280). The primary antibodies included anti-Ap-2α (#3215, Cell Signaling; the dilution ratio was 1:1000), anti-Elk-1 (#9182, Cell Signaling; the dilution ratio was 1:1000), anti-Sirpα (#13379, Cell Signaling; the dilution ratio was 1:1000) and anti-GAPDH (#5174, Cell Signaling; the dilution ratio was 1:2000) antibodies. Images were cropped for presentation.