Sc 271430

The immunohistochemical tissues of human and mouse were obtained from patients of the Department of Neurosurgery, Tongji Hospital, Huazhong University of Science and Technology (Wuhan, China) and mouse CDA model. Immunohistochemistry (IHC) primary antibody used in this study includes Goat anti-LRIG3 (1:50, R&D, AF3495), Rabbit anti-CD163 (1:100, Abcam, ab182422), Rabbit anti-IBA1 (1:2000, Abcam, ab178846), Rabbit anti-NETO2 (1:200, Sigma-Aldrich, HPA013180), Mouse anti-ARG1 (1:200, Santa Cruz, sc-271430), Rabbit anti-CD8 alpha (1:500, Abcam, ab209775) Rabbit anti-BAX (1:300, Boster, BA0412), and Rabbit anti-BCL2 (1:300, Boster, BA0315-1). The IOD scores of LRIG3, CD163, IBA1, ARG1 or CD8A in human GBM or mouse CDA models were quantified by software imageJ under ×400 magnified microscope with an Olympus Imaging System Microscope in the whole regions of each tumor specimen.

Total protein was extracted from myocardial tissue in the peripheral zone of MI and BMDMs, and the protein concentration was measured by a bicinchoninic acid assay kit. The sample loading amount was determined according to the sample concentration to ensure that the total protein loading amount of each sample was 40 μg. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis was performed, and the proteins were transferred to a polyvinylidene difluoride membrane. The membrane was incubated with primary antibodies against MCP-1 (Bioss, bs-34021R, 1:500), CCR2 (Biorbyt, orb378630, 1:1,000), Arg-1 (Santa Cruz, sc-271430, 1:500), MMP-2 (Abcam, ab92536, 1:1,000), MMP-9 (Abcam, ab76003, 1:1,000), and PPARγ (Abcam, ab209350, 1:1,000) overnight at 4°C and then incubated with HRP-conjugated goat anti-rabbit secondary antibody (ASPEN, AS1107, 1:10,000) at 37°C for 45 min; then, enhanced chemiluminescence detection was performed. Optical density was detected by the AlphaEaseFC software system.