Figure S4.
Subcellular domain-specific regulation of PS exposure and microglial contacts of spines of adult-born neurons. (A) Classification of spine morphology. Asterisks, mushroom; arrows, stubby; yellow arrowheads, thin; black arrowheads, filopodia. (B and C) Representative projection image of the EPL in the OB of adult Dcx-CreERT2; R26-tdTomato mice at 28 dpi (n = 8 mice), stained for DsRed (red), PS (white), and Iba1 (green). The boxed area in B is enlarged in C. Arrows indicate a PS+ tdTomato+ thin spine wrapped by Iba1+ microglial process. The two sequential z-planes (0, 0.5 µm) and a projection image (−1.0 to approximately +1.5 µm) of the boxed area in B are shown in C. (D) Representative projection image of the DG in the hippocampus of adult Dcx-CreERT2; R26-tdTomato mice at 28 dpi (n = 8 mice), stained for DsRed (red), PS (white), and Iba1 (green). Arrows indicate a PS+tdTomato+ spine wrapped by Iba1+ microglia. (E) Parameters for microglia–spine interaction (process distribution, touching, spreading and wrapping, and PS exposure). (F–I) Densities of microglial process distribution (F, n = 3 mice, pooled from two independent experiments), touching to spines (G, n = 4 mice, pooled from two independent experiments), spreading on and wrapping of spines (H, n = 4 mice, pooled from two independent experiments; F(3,12) = 30.7, P = 0.0000065, one-way ANOVA; basal versus proximal, P = 0.000037, basal versus apical, P = 0.000032, distal versus proximal, P = 0.00048, distal versus apical, P = 0.00040, Tukey–Kramer test), and proportion of PS+ spines (I, n = 3 mice, pooled from two independent experiments; F(3,8) = 16.7, P = 0.00083, one-way ANOVA; basal versus proximal, P = 0.0024, basal versus apical, P = 0.0019, distal versus proximal, P = 0.016, distal versus apical, P = 0.012, Tukey–Kramer test) in the basal, proximal, distal, and apical domains of dendrites of tdTomato+ adult-born granule cells in Dcx-CreERT2; R26-tdTomato mice at 28 dpi. EPL, external plexiform layer; ML, molecular layer. Scale bars in A–D, 2 µm. *, P < 0.05; ***, P < 0.005.

Subcellular domain-specific regulation of PS exposure and microglial contacts of spines of adult-born neurons. (A) Classification of spine morphology. Asterisks, mushroom; arrows, stubby; yellow arrowheads, thin; black arrowheads, filopodia. (B and C) Representative projection image of the EPL in the OB of adult Dcx-CreERT2; R26-tdTomato mice at 28 dpi (n = 8 mice), stained for DsRed (red), PS (white), and Iba1 (green). The boxed area in B is enlarged in C. Arrows indicate a PS+ tdTomato+ thin spine wrapped by Iba1+ microglial process. The two sequential z-planes (0, 0.5 µm) and a projection image (−1.0 to approximately +1.5 µm) of the boxed area in B are shown in C. (D) Representative projection image of the DG in the hippocampus of adult Dcx-CreERT2; R26-tdTomato mice at 28 dpi (n = 8 mice), stained for DsRed (red), PS (white), and Iba1 (green). Arrows indicate a PS+tdTomato+ spine wrapped by Iba1+ microglia. (E) Parameters for microglia–spine interaction (process distribution, touching, spreading and wrapping, and PS exposure). (F–I) Densities of microglial process distribution (F, n = 3 mice, pooled from two independent experiments), touching to spines (G, n = 4 mice, pooled from two independent experiments), spreading on and wrapping of spines (H, n = 4 mice, pooled from two independent experiments; F(3,12) = 30.7, P = 0.0000065, one-way ANOVA; basal versus proximal, P = 0.000037, basal versus apical, P = 0.000032, distal versus proximal, P = 0.00048, distal versus apical, P = 0.00040, Tukey–Kramer test), and proportion of PS+ spines (I, n = 3 mice, pooled from two independent experiments; F(3,8) = 16.7, P = 0.00083, one-way ANOVA; basal versus proximal, P = 0.0024, basal versus apical, P = 0.0019, distal versus proximal, P = 0.016, distal versus apical, P = 0.012, Tukey–Kramer test) in the basal, proximal, distal, and apical domains of dendrites of tdTomato+ adult-born granule cells in Dcx-CreERT2; R26-tdTomato mice at 28 dpi. EPL, external plexiform layer; ML, molecular layer. Scale bars in A–D, 2 µm. *, P < 0.05; ***, P < 0.005.

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