Journal: bioRxiv

Article Title: Persistent cortical excitatory neuron dysregulation in adult Chd8 haploinsufficient mice

doi: 10.1101/2025.06.04.657776

Figure Lengend Snippet: A) Barplot of DEG counts across bulk RNA-seq experiments (constitutive Chd8 +/- , Camk2a-CRE conditional Chd8 +/flox , and nuclear and synaptosomal preparations from the Chd8 +/- line. B) Chd8 is down-regulated in all but the nuclear preparation. C) LogFC concordance between constitutive and conditional datasets for constitutive model DEGs. D) LogFC concordance plots for nuclear and synaptosomal versus whole cortex for DEGs identified in nulear or synaptosomal fractions on left, reduced concordance between nuclear and synapsomal DEG logFC on right. E) Selected enriched GO terms for upregulated (left) and downregulated (right) genes across experiments. F) Downregulation of Dlg2 in synaptosomal RNA. GH) Western blot validating decreased synaptosomal PSD93 protein in Chd8 +/- cortex.

Article Snippet: Primary antibody PSD93 (1:50, DSHB, Clone N18/28, supernatant fraction) was diluted in 7.5 ml Intercept PBS blocking buffer with 0.1% Tween.

Techniques: RNA Sequencing, Western Blot

Journal: PLoS ONE

Article Title: Phosphoprotein Associated with Glycosphingolipid-Enriched Microdomains Differentially Modulates Src Kinase Activity in Brain Maturation

doi: 10.1371/journal.pone.0023978

Figure Lengend Snippet: A ) Whole brain lysates from WT mice of the indicated ages were Western blotted and stained for PSD93. B ) Brains of P1 wild type and Pag1 -/- mice and 3 month old WT/ Pag1 -/- littermate pairs were lysed at concentrations of 3 ml/g and 4.5 ml/g, respectively, in GEM-lysis buffer and subjected to sucrose-density centrifugation. Fractions were analyzed by Western blotting and staining for PSD93. Lipid raft fractions were identified by binding of cholera toxin subunit B (Ctx) binding to GM1-lipids in a dot blot from the fractions. C ) Densitometric quantification of the PSD93 signal in GEM-fractions as percentage of the total PSD93-signal. For both time points, the PSD93 signal in fractions 2-5 was expressed as a ratio to control. This most likely overestimates the lipid raft localized material in P1, as only fraction 2 and 3 contain rafts (n = 3 independent experiments for each time point) D ) Csk-immunoprecipitates from whole brain lysates of WT mice (ages as indicated), stained for PSD93 and Csk. Densitometric quantification of the relative amount of PSD93 bound to Csk in brains of P1 and 3 month old mice (n = 3 independent experiments) E ) Csk-immunoprecipitates from whole brain lysates of 3 month old WT and Pag1 -/- mice stained for PSD93 and Csk. Two exposures are given for the PSD93-signal in the left panel. Comparison of whole brain lysates of 3 month old WT and Pag1 -/- to P1. Given are the means +/- SEM, n.s. not significant, Student's t-test.

Article Snippet: Rabbit phosphor-specific anti-Src (pY418), recognizing both Y423 in mouse Src and Y419 in mouse Fyn, as well as anti-Src (pY529), recognizing both Y534 in mouse Src and Y530 in mouse Fyn due to the highly conserved sequence of these residues were purchased from BioSource, rabbit anti-Csk (C-20) and anti-Ctk from Santa Cruz, mouse-anti-Csk (clone 52) and mouse anti-CHK from BD BioSciences, mouse anti-Src (clone GD11) from Upstate (Lake Placid, NY), rabbit anti-PSD93 from Alomone Labs Ltd., rabbit anti-c-Src (Y215) from ECM Biosciences, and mouse anti-β-actin (clone AC-15) from Sigma.

Techniques: Western Blot, Staining, Lysis, Centrifugation, Binding Assay, Dot Blot