Fig. 2: Long-term exposure to HMF impeded newborn neuronal differentiation in the DG of mice.

a Schematic diagram showing the cell lineage-specific mapping markers across neurogenesis stages. b The experiment’s timeline for analyzing cell fate specification of aNSCs during HMF or GMF exposure. c Representative images of BrdU⁺ cells in the adult DG 1-, 2-, 4-weeks after BrdU injections. Scale bar = 200 μm. d Quantification of numbers of BrdU⁺ cells in GMF- and HMF-exposed mice 1-, 2-, 4-weeks after BrdU injections (GMF versus HMF, two-way ANOVA, F(1, 18) = 76.82, P < 0.0001). e Representative images of brain section stained with BrdU, GFAP, and S100β in the adult DG. Scale bar = 20 μm. f, g Quantification of numbers of BrdU⁺GFAP⁺S100β⁺ cells (f, GMF versus HMF, two-way ANOVA, F(1, 18) = 2.596, P = 0.1279) and BrdU⁺GFAP⁺S100β^– cells (g, GMF versus HMF, two-way ANOVA, F(1, 18) = 230.9, P < 0.0001) in GMF- and HMF-exposed mice 1-, 2-, 4-weeks after BrdU injections. h Representative images of brain section stained with BrdU, Ki67, and DCX in the adult DG. Scale bar = 20 μm. i, j Quantification of numbers of BrdU⁺Ki67⁺DCX^– cells (i, GMF versus HMF, two-way ANOVA, F(1, 18) = 88.54, P < 0.0001) and BrdU⁺Ki67⁺DCX⁺ (j, GMF versus HMF, two-way ANOVA, F(1, 18) = 88.54, P < 0.0001) cells in GMF- and HMF-exposed mice 1-, 2-, 4-weeks after BrdU injections. k Representative images of brain section stained with BrdU, DCX, and NeuN in the adult DG. Scale bar = 20 μm. l, m Quantification of numbers of BrdU⁺DCX⁺NeuN⁺ cells (l, GMF versus HMF, two-way ANOVA, F(1, 18) = 88.54, P < 0.0001) and BrdU⁺DCX^–NeuN⁺ cells (m, GMF versus HMF, two-way ANOVA, F(1, 18) = 16.93, P = 0.0007) in GMF- and HMF-exposed mice 1-, 2-, 4-weeks after BrdU injections. GMF, n = 4 mice, HMF, n = 4 mice. Data are presented as mean ± SEM. All data were analyzed by two-way ANOVA, and the two-tailed unpaired t test for two-group comparisons at each time point.