Anti cd3 fitc

For 15 min on ice, 10⁶ cells of SK-BR-3 (Her2⁺) (obtained from ATCC) or HPB-ALL (CD3⁺) (obtained from Cell resource center for biomedical research, Tohoku university; ID: TKG 0199) were incubated with 30 nM of IgG–Fab² in phosphate-buffered saline (PBS). An anti-human Fc FITC conjugate antibody (Sigma-Aldrich, MO USA) was added after washing the cells with PBS twice. PBS-suspended cells were used for the analysis using an RF-500 flow cytometry machine (Sysmex, Kobe, Japan) after washing the cells with PBS twice. The analyses used 100 μL of cell suspensions for each measurement. FCSalyzer software (Souceforge; https://sourceforge.net/projects/fcsalyzer/) was used to analyze the obtained data. As positive controls, trastuzumab (Chugai Pharmaceutical Co., Ltd, Tokyo, Japan) and anti-CD3 FITC (Proteintech Group, Inc., IL, USA) were used for SK-BR3 and HPB-ALL, respectively.

Atorvastatin (ATO, catalog number: BD18106) and cyclophosphamide (CTX, catalog number: BD122996) were purchased from Bidepharm (China). Pluronic F127 was purchased from Sigma-Aldrich (United States). ATP Assay Kit (catalog number: S0026) was purchased from Beyotime biotechnology (China). Anti-CD16/32 (catalog number: 65080-1-L), anti-CD3-FITC (catalog number: FITC-65077), and anti-CD8a-APC (catalog number: APC-65069) were purchased from Proteintech (China). Anti-CD274 (PD-L1)-PE (catalog number: 124308) was purchased from Biolegend (United States). Anti-calreticulin primary antibody (catalog number: ab271865) was purchased from Abcam (United States). HMGB1 rabbit monoclonal antibody (catalog number: AG2167), β-actin mouse monoclonal antibody (catalog number: AF0003), Alexa Fluor 488-labeled goat anti-rabbit IgG (H + L) (catalog number: A0423), HRP-conjugated goat anti-mouse IgG (H + L) (catalog number: A0216), and HRP-conjugated goat anti-rabbit IgG (H + L) (catalog number: A0208) were purchased from Beyotime biotechnology (China).