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. 2021 Jan 7;11:611133. doi: 10.3389/fimmu.2020.611133

Figure 2.

Figure 2

TGF-β and IL-6 are responsible for activated platelet-mediated induction of CD16 on monocytes. (A) Quantitative RT-PCR analysis of TGF-β, IL-10, and IL-6 in monocytes co-cultured with activated platelets at the indicated time points. mRNA expression of each gene was normalized to that in monocytes incubated without platelets at the same time points (n = 5 or 6). (B) The amount of TGF-β, IL-10, and IL-6 in supernatants of monocytes co-cultured with or without ADP-activated platelets (n = 6 or 7) or unstimulated platelets (n = 3) were quantified (ELISA). (C, D) Representative contour plot of CD16 expression on monocytes co-cultured with activated platelets in the presence of anti-TGF-β (10 μg/ml), anti-IL-10 (10 μg/ml), or anti-IL-6 (10 μg/ml) neutralizing antibodies (left panels of C and D). Purified CD14+CD16- monocytes were incubated with each neutralizing antibody for 30 min, followed by addition of ADP-activated platelets into the culture. Frequencies (%) of CD14+CD16+ monocytes under the indicated conditions were analyzed using flow cytometry (right panels of C and D) (n = 4 or 5). Bars show the mean ± S.E.M. *p < 0.05, **p < 0.01, ***p < 0.005, and ****p < 0.001 by two-tailed paired t-test. NS, not significant.