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. 2023 Sep 21;14:1209249. doi: 10.3389/fimmu.2023.1209249

Figure 7.

Figure 7

TRAIL increases the expression of classical M1 markers in primary human M1 macrophages at mRNA and protein levels. (A) Macrophages were pre-stimulated with 200 ng/ml TRAIL for 2 hours and then polarized into M1 with 100 ng/ml LPS and 20 ng/ml IFNγ for 6 hours. Control group was stimulated only with LPS and IFNγ for 6 hours. Expression of M1 markers was analyzed by qPCR. (B) Macrophages were pre-stimulated with 200 ng/ml TRAIL for 6 hours and then polarized into M1 with 100 ng/ml LPS and 20 ng/ml IFNγ for 12 hours. Control groups were left unstimulated or stimulated only with LPS and IFNγ for 12 hours. Expression of CD86, HLA-DR alpha, CD64, CXCL10, TNF, and CXCL1 was analyzed by flow cytometry, and representative plots are included. Production of IL-1β, TNF, and IL-12 p70 was analyzed by ELISA. Data shown are mean ± SEM or median with interquartile range pooled from two or more independent experiments [(A) n=8-13, (B) n= 6-16]. Statistical analyses were performed with a two-tailed paired Student’s t-test, Wilcoxon matched-pairs signed-rank test, One-way ANOVA with Sidak’s multiple comparisons post-hoc test, or Friedman with Dunn’s multiple comparisons post-hoc test between untreated and M1 macrophages, M1 and TRAIL-treated M1 macrophages, *P<0.05, **P<0.01, ***P<0.001.