Alexa fluor 488 anti mouse cd86

Abemaciclib was purchased from MedChemExpress Co. Ltd. IMD‐0354 was purchased from Selleckchem Co. Ltd. MAL‐PEG₂₀₀₀‐NHS was purchased from Beijing Kaizheng Biotech Co. Ltd (Beijing, China). Poly (L‐lysine) (PLL, M_w = 3–7 w), 2,3‐Dimethylmaleic Anhydride (DMA), Cis‐aconitic acid anhydride (CA), 3,4,5,6‐Tetrahydrophthalic anhydride (TDA) and succinic anhydride (SA) were purchased from Sigma‐Aldrich Co. Ltd (American). NGR peptide (sequence: GCNGRCGC) was obtained from Shanghai Apeptide Co. Ltd (Shanghai, China). Polyamindoamine (PAMAM‐G5, M_w = 28 826) was purchased from CY dendrimer technology Co. Ltd (Weihai, China). Cell Cycle and Apoptosis Analysis Kit was the product of Beyotime Biotechnology Co. Ltd (Shanghai, China). Alexa Fluor 647 anti‐mouse F4/80, Brilliant Violet 421 anti‐mouse CD206, PerCP/Cy5.5 anti‐mouse F4/80, Alexa Fluor 488 anti‐mouse CD86, APC anti‐mouse CD206, APC anti‐mouse CD3, FITC anti‐mouse CD4, PE anti‐mouse CD8, PE anti‐mouse CD25, and Alexa Fluor 488 anti‐mouse Foxp3 were purchased from Biolegend. ELISA kits were obtained from Dakewei Co. Ltd (Nanjing, China). All other reagents and solvents were obtained from Sinopharm Co., Ltd (Shanghai, China) and Sigma‐Aldrich Co., Ltd (Shanghai, China).

To analyze M1-like macrophages, BMDMs were stained with PerCP/Cyanine5.5 anti-mouse/human CD11b (101228; BioLegend), APC anti-mouse F4/80 (123116; BioLegend), and Alexa Fluor 488® anti-mouse CD86 (105017; BioLegend). For intracellular iNOS staining, BMDMs after Fc-blocking were stained with PerCP/Cyanine5.5 anti-mouse/human CD11b (101228; BioLegend) and APC anti-mouse F4/80 (123116; BioLegend) at 4°C for 30 min, fixed and permeabilized with BD Cytofix/Cytoperm Kit (554714; BD Biosciences) at 4°C for 20 min. After washing with Perm/Wash buffer (BD Biosciences), the cells were stained with Alexa Fluor 488 anti-iNOS (53-5920-80; Invitrogen) at 4°C for 30 min. The stained cells were resuspended in 2% paraformaldehyde and analyzed using the NovoCyte 2060 R (ACEA Biosciences, USA), using FlowJo software ver 10.7.2 (BD Biosciences)