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. 2022 Jan 20;13:773261. doi: 10.3389/fimmu.2022.773261

Figure 2.

Figure 2

Acetate increases nitric oxide production via IL-1β. (A) Nitrite concentration assessed by Griess assay from supernatant of MPI cells pre-treated with conditioned medium, then stimulated or not with S. pneumoniae. Conditioned medium is the supernatant of MPI cells stimulated with S. pneumoniae in the presence or absence of acetate during 24 h. (B) ELISA of IL-1β from supernatant of MPI cells pre-treated or not with acetate and stimulated or not with S. pneumoniae during 24 h. (C, D) Kinetics from MPI cells pre-treated with acetate and then stimulated with S. pneumoniae. Cellular lysate was used for Il1b and Nos2 mRNA quantification by RT-PCR, and supernatants for IL-1β secretion by ELISA and nitrite by Griess assay. For RT-PCR fold increase was calculated over time point 0 (Vh). (E) Nitrite concentration assessed by Griess assay from supernatant of MPI cells: Pre-treated or not with IL-1β and stimulated or not with S. pneumoniae during 24 h (E) left panel). Pre-treated or not with acetate and/or anti-IL-1β antibody and stimulated of not with S. pneumoniae for 48 h (E) right panel). Bars showing the mean and error showing the SEM of triplicates. Results are representative of three independent experiments. Statistical analysis was done using Two-way ANOVA corrected with Sidak’s multiple comparisons test (**p < 0.01, ***p < 0.001, ****p < 0.0001).