Additional file 1: of Purinergic targeting enhances immunotherapy of CD73+ solid tumors with piggyBac-engineered chimeric antigen receptor natural killer cells

Figure S1. (A) Scheme of the synthesis of PBAE 447. (B) 1 H NMR spectra of PBAE 447 polymer in DMSO-d6. Figure S2. Cell viability of NK-92 cells after incubation with PBAE 447 polymer at various concentrations for 52 h. Figure S3. Flow cytometric analyses of NKG2D expression in NK-92 cells. NK-92 cells were stained with FITC-NKG2D and positive staining was analyzed. Figure S4. The mean fluorescence intensity (MFI) of NKG2D expression following 5 days’ continuous culture of NKG2D.CAR-NK-92 cells. Figure S5. (A) Cell viability of NK-92 cells after incubation with TGF-β1 for 24 h. (B) The effect of TGF-β1 treatment on NKG2D expression by NK-92 cells. Figure S6. (A) Percentage of NK cells producing IFN-γ intracellularly measured by flow cytometry. (B) Degree of degranulation of NK cells expressed as % CD107a+ cells. Figure S7. Cell viability of NK-92 cells after incubation with adenosine (ADO) at various concentrations for 24 h. Figure S8. Lytic activity of NK-92 cells against (A) GBM43, (B) GBM10, (C) A549 or (D) PC3 cells, in the presence of anti-CD73 antibody (10 μg/mL) and adenosine deaminase inhibitor (ADAi) EHNA (30 μM), respectively. Figure S9. CD73 expression on (A) A549 and (B) GBM10 cells after treatment with TGF-β1 for 24 h. Figure S10. (A) CD73 expression on K562 cells. (B) Lytic activity of NK-92 and piggyBac-NKG2D.CAR-NK-92 cells against CD73- K562 cells. (DOCX 914 kb