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. Author manuscript; available in PMC: 2020 Jul 27.
Published in final edited form as: Nat Microbiol. 2020 Jan 27;5(3):511–524. doi: 10.1038/s41564-019-0649-5

Fig. 4. F. PB1 releases SCFAs that have anti-proliferative activity.

Fig. 4

a,c, Cell proliferation assay on mouse CRC cell lines treated or not (NT) with acetate (Ac), propionate (Prop) and butyrate (But) either alone and in combination (MIX) (a) or with culture broth fermented by F. PB1 (SUP) (c). t0 is the signal from cells at the time of stimulation. Two independent experiments were performed with consistent results. Data from one representative experiment (n = 6 biologically independent samples). P values were determined by one-way ANOVA using Bonferroni post-test. b, Quantification of L-lactate and SCFAs in broth fermented by F. PB1 (SUP) by UPLC-MS. Data from six independent experiments. d, Representative Western blots from two to three independent experiments showing the effect on H3K27 acetylation, PP2B-A and NFATc3 expression in mouse cell lines treated or not (NT) with broths fermented by F. PB1 (SUP) or not fermented (Veh). Vinculin and actin were used as loading controls. Densitometric analysis is reported in Extended Data Fig. 5b. e,f, In vitro stimulation of CT26 cells with untreated broth fermented by F. PB1 (SUP) or one depleted of SCFAs by evaporation (SUP evap). Untreated broth not fermented (Veh) or evaporated (Veh evap) used as controls. e, Representative Western blots from two independent experiments showing the effect of SUP and SUP evap on H3K27 acetylation and NFATc3 expression. Vinculin was used as loading control. Densitometric analysis is reported in Extended Data Fig. 5c. f, Quantification of SCFAs and L-lactate by UPLC-MS. n = 3 (SUP evap) or 6 (SUP) biologically independent experiments. P values were determined by two-tailed unpaired Mann-Whitney test. a,b,c,f, Data are presented as means ± s.d.