Journal: Nature neuroscience

Article Title: Chd8 mediates cortical neurogenesis via transcriptional regulation of cell cycle and Wnt signaling.

doi: 10.1038/nn.4400

Figure Lengend Snippet: Figure 6 Chd8 Knockdown in upper cortical layer neurons results in behavioral deficits, which can be rescued via induction of Wnt signaling. (a) Chd8- knockdown mice show reduced number of entries to the light chamber in light–dark chamber assay (one-way ANOVA followed by Bonferroni’s multiple comparison test; DF = 2, F = 18.70, P < 0.0001; Control; n = 17, Chd8 sh2; n = 8, Chd8 sh2 + S/A-β-cat; n = 10). n, number of animals used. (b) Chd8-knockdown mice spent less time in the light chamber (one-way ANOVA followed by Bonferroni’s multiple comparison test; DF = 2, F = 9.521, P = 0.0006; Control; n = 17, Chd8 sh2; n = 8, Chd8 sh2 + S/A-β-cat; n = 10). n, number of animals used. (c) Number of entries to the open arms in elevated plus maze assay was significantly reduced in Chd8-knockdown mice (one-way ANOVA followed by Bonferroni’s multiple comparison test; DF = 2, F = 23.27, P < 0.0001; Control; n = 16, Chd8 sh2; n = 12, Chd8 sh2 + S/A-β-cat; n = 10). n, number of animals used. (d) Chd8 knockdown mice spent significantly less time in the light chamber (one-way ANOVA followed by Bonferroni’s multiple comparison test; DF = 2, F = 16.54, P < 0.0001; Control; n = 16, Chd8 sh2; n = 12, Chd8 sh2 + S/A-β-cat; n = 10). n, number of animals used. (e) In the 3-chamber social interaction assay, Chd8- knockdown animals did not spend significantly more time in the chamber with the novel social animal (Stranger) relative to the chamber with the empty wire cage (Empty) compared to control or to the Chd8 sh2 + S/A-β-cat group (One-way ANOVA followed by Bonferroni’s multiple comparison test; DF = 2, F = 43.09, P < 0.0001 for Control; DF = 2, F = 3.167, P = 0.0552 for Chd8 sh2; DF = 2, F = 19.81, P < 0.0001 for Chd8 sh2 + S/A-β-cat; Control; n = 13, Chd8 sh2; n = 12, Chd8 sh2 + S/A-β-cat; n = 10). n, number of animals used. (f) Social interaction in close proximity (within 5 cm of the wire cages). Chd8-knockdown animals did not show preference for either the Stranger or the Empty cage (Two-tailed unpaired student’s t-test; t = 6.942, DF = 10, P <0 .0001, n = 6 for Control; t = 0.4274, DF = 14, P = 0.6756 n = 8 for Chd8 sh2; t = 9.930, DF = 18, P < 0.0001, n = 10 for Chd8 sh2 + S/A-β-cat). (g) Images of upper cortical neurons from 5-month-old mouse brains transfected at E15 with either control shRNA or Chd8 shRNA and membrane-bound GFP. Scale bars: 20 µm. (h) Chd8 knockdown reduced the complexity of dendritic arborization assessed via Sholl analysis (two-way ANOVA followed by Bonferroni’s multiple comparison test; DF = 2, sum-of-squares = 4,226, mean square = 2,113, F = 269.5, P < 0.0001 for genotype factor; Control; n = 24 cells from 5 animals, Chd8 sh2; n = 52 cells from 7 animals, Chd8 sh2 + S/A-β-cat; n = 17 cells from 5 animals). *P < 0.05; **P < 0.01; ***P < 0.001; n.s., nonsignificant. Results are presented as mean ± s.e.m.

Article Snippet: The following primary antibodies were used in this study: rabbit anti-CHD8 antibody (A301-224A, Bethyl Laboratories; ChIP), rabbit anti-Chd8 antibody (noncommercialized, Cell Signaling; 1:1,000 dilution; WB), chicken anti-GFP antibody (GFP-1020, Aves Labs; 1:1,000 dilution; IHC), mouse anti-BrdU antibody (M0744, Clone Bu20a, DakoCytomation; 1:500; IHC), rabbit anti-Ki67 antibody (Clone SP6, Lab Vision/Thermo Scientific; 1:500 dilution; IHC), rabbit anti-phospho-histone H3 (Ser10) antibody (06-570, Millipore; 1:500 dilution; IHC), mouse anti-Tuj1 antibody (MMS-435P, Covance; 1:500 dilution; IHC) and mouse anti-EZH2 antibody (AC22, Active Motif; 1:500 dilution, WB).

Techniques: Knockdown, Boyden Chamber Assay, Comparison, Control, EPM Assay, Two Tailed Test, Transfection, shRNA, Membrane

Journal: Neuron

Article Title: Polyserine-mediated targeting of FAF2/UBXD8 ameliorates tau aggregation

doi: 10.1016/j.neuron.2025.08.002

Figure Lengend Snippet: ( A ) Immunostaining of DAPI (blue), phosphorylated tau (AH36) (green) and NeuN (magenta) in hippocampi of WT or PS19 animals injected with AAV9-GFP, AAV9-FAF2 or AAV9-FAF2 PS . ( B ) Quantification of AH36+ hippocampal area as in (A). One-way ANOVA and Tukey’s MCT. n=3 sections per animal averaged for n=10 animals per group. ( C ) Volcano plot of protein expression by Tandem mass tag pro (TMTpro) mass spectrometry in brain extracts from AAV9-FAF2 animals compared to AAV9-GFP at 6-months-old. Shaded areas represent fold change >2 and p-value >0.01. ( D ) Volcano plot of protein expression by TMTpro mass spectrometry in brain extracts from AAV9-FAF2 PS animals compared to AAV9-GFP at 6-months-old. Shaded areas represent fold change >2 and p-value above 0.01. ( E ) Percent of time spent on closed arms during elevated plus maze assay for 10-month-old animals as in (A). Boxplot whiskers by Tukey’s method. One-way ANOVA. WT+GFP(n=16), PS19+GFP(n=18), PS19+FAF2(n=11), PS19+FAF2 PS (n=12). ( F ) Percent freezing during training period of fear conditioning assay for animals as in (E). Grey boxes denote cued stimulus (tone) and red line denotes aversive stimulus (foot shock). WT+GFP(n=18), PS19+GFP(n=19), PS19+FAF2(n=11), PS19+FAF2 PS (n=11). ( G ) Average percent of time spent freezing during periods of cued stimulus for animals as in (F). Boxplot whiskers by Tukey’s method. One-way ANOVA and Tukey’s MCT. WT+GFP(n=18), PS19+GFP(n=19), PS19+FAF2(n=11), PS19+FAF2 PS (n=11). ( I ) Average percent of time spent freezing during contextual testing for animals in (F). Boxplot whiskers by Tukey’s method. One-way ANOVA and Tukey’s MCT. WT+GFP(n=18), PS19+GFP(n=19), PS19+FAF2(n=11), PS19+FAF2 PS (n=11).

Article Snippet: Rabbit anti-p-tau (AH36) , StressMarq Biosciences , Cat#SMC-601; RRID:AB_2820300.

Techniques: Immunostaining, Injection, Expressing, Mass Spectrometry, EPM Assay