Figure 1.

C24D induced re-activation of leukocytes, resulting in killing of TNBC cells: a. Representative pictures of tumor cell density in co-cultures of MDA-MB-231 cells with PBMCs from healthy female donors, documented by inverted microscope (Bars = 200 μm) ± C24D (10 μg/ml) or scrambled C24D (S-C24D, 10 μg/ml) as control. b. Tumor cell killing by FACS analysis, using AnnaexinV/PI (N = 14). c. Representative FACS dot blot (experiment #11) of AnnaexinV/PI gated and stained tumor (CD45 negative) cells treated with 10 μg/ml C24D for 4–6 days. d. Percentage of CD4⁺/CD69⁺ and CD4⁺/CD57⁺ cells in co-cultures of PBMC with MBA-MB-231 cells, ± C24D, by FACS analysis. e. Percentage of CD8+/CD69+, CD8+/CD56+ (T/NK cells) and CD8+/CD57+, in the co-cultures, by FACS analysis. f. Percentage of CD56+/CD69+ and CD56+/CD57+, in the co-cultures, by FACS analysis. g. Percentage of CD14+/CD137+ in the co-cultures, by FACS analysis. Analysis of the results presented as Mean ± SD. h. Representative dot blot FACS analysis showing double staining of CD4, CD8 or CD56 gated cells with either CD69 or CD57, comparing effect of C24D versus control. i. Effect of C24D on IFNγ secretion (pg/ml) in supernatants of PBMCs alone or PBMCs co-cultured with MDA-MB-231 (n = 8). j. Effect of C24D on secretion of IL-2 (pg/ml) (n = 7). k. Effect of C24D on secretion of TNFα (pg/ml) (n = 7). l. Effect of C24D on secretion of IL-1β (pg/ml). Analysis of results presented as Mean ± SD