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. 2020 Oct 6;2(2):fcaa165. doi: 10.1093/braincomms/fcaa165

Figure 7.

Figure 7

Short-chain fatty acids trigger neuronal differentiation of NSCs in a ROS- and p-ERK1/2-dependent manner. Mouse NSCs were treated with 1 mM of propionate (n = 3–7) and butyrate (n = 3–7) and 0.5 μM NAC for 24 h in self-renewal conditions. Cells were collected for qRT-PCR and Western blot analysis, as described in Materials and methods section. Effect of SCFAs on βIII-tubulin and NeuN (A), and Sox2 (B) mRNA levels. Hprt was used as loading control. A time-dependent assay was performed to evaluate the effect of propionate and butyrate on ERK1/2 phosphorylation. As described in Materials and methods section, NSCs were transfected with a MEK over-expression plasmid to evaluate the impact of ERK1/2 activation on SCFA-mediated changes in NSC fate. (C) Representative immunoblots (top) of p-ERK1/2 protein levels and respective densitometry analysis (bottom). β-Actin was used as loading control. (D) Representative immunoblots showing the efficiency of MEK transfection in NSCs. (E) Effect of ERK1/2 activation on SCFA-mediated changes in βIII-tubulin and NeuN, and Ki67 mRNA levels (F). Hprt was used as loading control. Data are expressed as fold change over control. Data represent mean values ± SEM for at least three independent experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 compared to untreated cells.