Fig. 5

Efficacy of the ADSC secretome to effector function of NK-92 cells when each immunomodulatory candidate within the ADSC secretome was blocked by neutralizing antibody. A to C NK-92 cells stimulated with IL-2 (5 ng/ml) were incubated for 48 h with the ADSC secretome (150 μg/ml) and neutralizing antibodies (1 μg/ml) to each candidate to examine recovery of cellular activity. Unstimulated NK cells were cultured for negative control. A The concentrations of IFN-γ and IL-10 in the culture supernatants was measured by ELISA. Error bars represent mean ± SEM of five independent experiments. Statistical tests were determined by Student’s paired two-tailed t-test; *P < 0.05. Anti-ANXA1, anti-annexin A1 antibody; anti-LGALS3BP, anti-galectin-3-binding protein antibody. B After incubation, NK-92 cells were co-cultured with K562 target cells for 4 h at the indicated E:T ratios, followed by measurement of cytotoxicity of NK-92 cells against K562 cells. C NK-92 cells were stained with PE-conjugated CD107a antibodies and co-cultured with K562 cells for 4 h at the indicated E:T ratios to measure the level of exocytosis of NK cells toward target cells. Degranulation marker CD107a was assessed using flow cytometry as shown dot blot (left) and frequency of CD107a⁺ NK cells (right). Representative data were shown