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. 2018 Jul 9;9:1569. doi: 10.3389/fimmu.2018.01569

Figure 3.

Figure 3

Trypanosoma cruzi-infected NIH-3T3 fibroblasts have characteristics of cellular senescence. NIH-3T3 fibroblasts were infected overnight with culture-derived T. cruzi trypomastigotes (MOI 5:1). (A) Evaluation of soluble SA-β-gal activity at pH 6.0 in cell lysates of T. cruzi-infected and non-infected fibroblasts 3 days post-infection. (B,C) Measurement of senescence-associated secretory phenotype (SASP) cytokines IL-6, TNF-α and IL-1β and chemokine MCP-1, as well as SASP non-related IL-10, in the 3 days post-infection culture supernatants by ELISA. (D) Evaluation of nitric oxide release by T. cruzi-infected and non-infected fibroblasts in culture supernatants 3 days post-infection by Griess reaction. Data are presented as the mean ± SE of at least six biological replicates and analyzed by (A) non-parametric Mann–Whitney test, and (B–D) unpaired, two-tailed Student’s t-test, *p < 0.05; **p < 0.01; ***p < 0.001; NS, statistically not significant, p > 0.05. Data are representative of at least three independent experiments with similar results. Abbreviations: MOI, multiplicity of infection; Ctrl, control; SA-β-gal, senescence-associated β-galactosidase.