Fitc labeled anti cd86 antibody

JQ1 (≥98% purity, CAS No. 1268524-70-4) and Oridonin (>98% purity, CAS No. 28957-04-2) were purchased from Meilunbio (Dalian, China). Tumor-penetrating peptide iRGD (internalizing RGD, ≥98% purity, CRGDKGPDC) was constructed by ChinaPeptides (Shanghai, China). PSS (20 kDa) was provided by Ocean University of China (Qingdao, China). ssPBAE was synthesized according to our previous report.31 (link) 1-(3-dimethylaminoprophyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl) and N-hydroxysuccinimide (NHS) were provided by J&K Scientific (Beijing, China). IR780 iodide, polyvinylpyrrolidone (PVP, K-30), thiazolyl blue tetrazolium bromide (MTT), 4ʹ6-diamidino-2’-phenylindole dihydrochloride (DAPI) and 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) were obtained from Sigma Aldrich (St Louis, USA). Singlet oxygen sensor green (SOSG) and LIVE/DEAD cell staining kit were purchased from Thermo Fisher Scientific (Hudson, NH, USA). L-Lactic acid and lipopolysaccharides (LPS) were obtained from Solarbio (Beijing, China). Anti-calreticulin/AF488 antibody was purchased from Biosynthesis Biotechnology (Beijing, China). Antibodies against PD-L1, GAPDH and HRP-conjugated antibodies were provided by Bioss Antibodies (Boston, MA, USA). The FITC-labeled anti-CD86 antibody, APC-labeled anti-CD80 antibody and antibody against Ki67 were obtained from Abcam (Cambridge, UK).

We measured the expressions of costimulatory molecules CD80 and CD86 of BMDCs to evaluate the maturation of BMDCs in vitro. Briefly, BMDCs (1.0 × 10⁶ cells/mL) were cultured with different concentrations of lactic acid (15 mM and 25 mM), as well as lipopolysaccharide (LPS, 36 µg/mL) as a positive control, respectively. After 24-hour incubation, BMDCs were stained with FITC-labeled anti-CD86 antibody (Abcam, Cambridge, UK) and APC-labeled anti-CD80 antibody (Abcam, Cambridge, UK), and subsequently analyzed by a flow cytometer. The percentage of cells staining positive for each surface protein was recorded.