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. 2022 Jan 20;17(2):259–275. doi: 10.1016/j.stemcr.2021.12.010

Figure 5.

Figure 5

ABT-263 administration enhances neurogenesis and NPCs in 12 month old mice

(A–N) 12 month old mice were injected with ABT-263 or vehicle and the DG was analyzed 5 days later. In (H and M) mice were also injected with BrdU 4 days post-ABT-263 and analyzed 24 h (M) or 30 days (H) later. (A and B) DG sections were stained for SA-β-Gal (A, black, arrows), and total SA-β-Gal-positive SGZ cells were quantified (B). Dashed lines indicate the SGZ/hilus border. (C) DG sections were stained for SA-β-Gal and immunostained for SOX2, S100β, or IBA1 (as in Figures 2C–2E), and total SA-β-Gal-positive SGZ cells co-expressing each of these markers were quantified. Also quantified were total SA-β-Gal-positive cells. (D and E) DG sections were immunostained for SOX2 (D, red, arrowheads) and IBA1 (D, green, arrows), and total IBA1-positive SGZ cells were quantified (E). The dashed line indicates the hilus/SGZ border. (F) qPCR for Il6 and Mmp2 mRNAs in total DG mRNA. Values were normalized to Gapdh mRNA levels in the same samples and expressed as a fold change in ABT-263 versus vehicle-treated samples. (G) Quantification of immunostained sections for total DCX-positive cells in the DG. (H) Quantification of total BrdU-positive, NeuN-positive cells in the DG of ABT-263 or vehicle-treated mice injected with BrdU and analyzed 30 days later. (I and J) DG sections were immunostained for SOX2 (I, green, arrows), and total SGZ SOX2-positive cells were quantified (J). (K) Quantification of immunostained sections for total GFAP-positive, SOX2-positive SGZ cells. (L and M) DG sections from mice injected with ABT-263/vehicle and then BrdU were immunostained 24 h post-BrdU to identify (L, green, arrows) and quantify (M) total BrdU-positive SGZ cells. (N) Quantification of immunostained DG sections for GFAP-positive, KI67-positive SGZ cells, expressed as a percentage of total GFAP-positive, SOX2-positive SGZ cells as determined in (K).

(O and P) 12 month old mice were injected ICV with 50 ng of ABT-263 or vehicle, and coronal DG sections were analyzed 5 days later for total SA-β-Gal-positive SGZ cells (O) or total DCX-positive DG cells (P). In all cases, error bars indicate SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. n = 3 mice/condition in all cases, except for (C), where n = 5 mice/condition, and (F), where n = 6 mice/condition. Scale bars: 50 μm (A, I), 25 μm (D), 20 μm (L). In (I and L) sections were counterstained with Hoechst 33258 (blue).