Journal: International Journal of Molecular Sciences

Article Title: FOXG1 Contributes Adult Hippocampal Neurogenesis in Mice

doi: 10.3390/ijms232314979

Figure Lengend Snippet: FOXG1 increases the amount of neuronal stem cells (aNSCs) in adult hippocampal DG of the Foxg1 genotype mice and promotes qNSCs giving rise to activated aNSCs. ( A ) Expression patterns of GFAP-expressing qNSCs in the hippocampal DG of Foxg1 genotype mice were assessed with immunofluorescence. Red = GFAP; Green = EGFP-labeled FOXG1; Blue = DAPI. Scale bars = 100 μm. Arrow heads: newly generated qNSCs with FOXG1 activation. ( D ) Levels of total qNSCs in adult hippocampus are showed as %GFAP + /DAPI + cells. ( E ) Levels of endogenous qNSCs in adult hippocampus are showed as %GFP − GFAP + /DAPI + cells. ( B ) Expression patterns and ( F ) qualification of total qNSCs in adult hippocampus were studied with immunohischemisitry using an antibody against GFAP. Scale bars = 100 μm. ( C ) Hippocampal lysates of the two genotype mice were immunoblotted using an antibody against GFAP. ( G ) Qualification of GFAP expression is illustrated by bar graph. GAPDH was used as the loading control. ( H ) Expression patterns of proliferated qNSCs in the hippocampal DG of Foxg1 genotype mice were assessed with immunofluorescence. Red = GFAP; Green = Proliferating Cell Nuclear Antigen (PCNA); Blue = DAPI. Scale bars = 100 μm. Arrow heads: proliferated qNSCs. ( J ) Levels of qNSCs in adult hippocampus are showed as %GFAP + /DAPI + cells. ( K ) Levels of proliferated qNSCs in adult hippocampus are showed as %GFAP + PCNA + /DAPI + cells. ( I ) Expression patterns of activated aNSCs in the hippocampal DG of Foxg1 genotype mice were assessed with immunofluorescence. Red = brain lipid binding protein (BLBP); Green = GFAP; Blue = DAPI. Scale bars = 100 μm. Arrow heads: activated aNSCs. ( L ) Levels of aNSCs in adult hippocampus are showed as %GFAP + /DAPI + cells. ( M ) Levels of activated aNSCs in adult hippocampus are showed as %GFAP + BLBP + /DAPI + cells. Values are expressed as means ± S. E. M. n = 6/group. Significant levels set at *** p < 0.001 noted difference between Foxg1 fl/fl and Foxg1 fl/fl -CreAAV animals. ns—not significant.

Article Snippet: To visualize protein expression, sections were first incubated with primary antibodies, including rabbit anti-GFAP (1:200, Wanleibio, cat # WL0836, Shenyang, China), mouse anti-GFAP (1:200, Proteintech, cat# 60190-1-lg, Rosemont, USA), EGFP (1:200, Beyotime, cat # AG281, Shanghai, China), brain lipid binding protein (BLBP) (1:200, Proteintech, cat # 51010-1-AP, Rosemont, USA), Proliferating Cell Nuclear Antigen (PCNA) (1:250, Proteintech, cat # 10205-2-AP, Rosemont, USA), Tbr2 (1:200, Bioss, cat # bs-11331R, Beijing, China), Oligo2 (1:200, Bioss, cat # bs-11194R, Beijing, China), NeuN (1:1200, Bioss, cat # bs-10394R, Beijing, China) and Dcx (1:200, Proteintech, cat # 13925-1-AP, Rosemont, USA).

Techniques: Expressing, Immunofluorescence, Labeling, Generated, Activation Assay, Control, Binding Assay