darpp32 antibody  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Oxidative Stress-Induced Microglial CD22 Upregulation Impairs Phagocytosis and Exacerbates Huntington’s Disease

doi: 10.64898/2026.03.11.710967

Figure Lengend Snippet: (a ) HD mouse model (R6/2) was crossed with CD22-knockout (KO) mice to evaluate the biological impact of CD22 in vivo. ( b ) Rotarod performance was assessed from 7 to 12 weeks of age. Knockout of CD22 significantly improved the rotarod performance of the HD mice. Data are presented as mean ± SEM (n = 9-12 animals per group). *Specific comparison between WT and HD mice in each condition; #Specific comparison between CD22-WT and CD22-KO in each condition by two-way ANOVA. Immunofluorescence staining of brain sections was performed to detect mHTT aggregates (green, EM48) ( c ) and the expression of the medium spiny neuron marker (green, Darpp32) ( e ) in the neurons (red, NeuN) of HD mice. ( d ) Quantification of (c). ( f ) Quantification of (e). Knockout of CD22 reduces the area and intensity of mHTT aggregates and the loss of Darpp32 in HD mice. Data are presented as mean ± SEM (n = 6-8 animals per group). * p < 0.05, ** or ## p < 0.01, *** p < 0.001 and **** p < 0.0001 by unpaired t -test. Scale bar = 10 µm.

Article Snippet: The primary antibodies included NeuN (Merck Millipore; ABN78), mEM48 (Merck Millipore; MAB5374), Darpp32 (Santa Cruz; sc-271111), Iba1 (FUJIFILM Wako Pure Chemical Corporation; 019-19741), and CD68 (Bio-Rad; MCA1957).

Techniques: Knock-Out, In Vivo, Comparison, Immunofluorescence, Staining, Expressing, Marker

Journal: bioRxiv

Article Title: Large-scale Perturbation of Systems Biology-Derived Genes Reveals Modifiers of HD- associated Transcriptomic Networks and Pathology

doi: 10.64898/2026.03.02.709091

Figure Lengend Snippet: (A) Boxplot of log-transformed normalized expression of Foxp1 in the 4 genotypes. Blue points show individual expression values, boxes show the median and interquartile range, notches indicate median confidence interval. (B) Enrichment of genes DE in Foxp1 KO-het vs. WT and Q140/Foxp1 KO-het vs. Q140 in selected terms that include BioSystems pathways, cell type markers and genes DE in studies of HD models. For clarity, the enrichment p-values are limited to 10-50. (C) Representative images of Foxp1 staining in striata of 6m Q140/Foxp1 KO-het cohort. Coronal striatal sections (30µm thickness). (D) Foxp1 co-localized with nuclear inclusions in the striatum of Q140 mice. (E) Foxp1 intensity in striata of 6m Q140/Foxp1 KO-het cohort (n=4 per genotype). For all bar graphs in this figure, data are represented as mean ± SEM. One-way ANOVA with Tukey’s HSD tests: * p<0.05, ** p<0.01, *** p<0.001, ****p<0.0001. (F) Representative images of EM48-stained mHtt aggregates in striata of 6m Q140/Foxp1 KO-het cohort. Coronal striatal sections (30µm thickness) were double stained with anti-Darpp32 (amber) and anti-mHtt (EM48, green) antibodies. (G) Darpp32 intensity in striata of 6m Q140/Foxp1 KO-het cohort (n=3 per genotype). (H) Representative Nissl-stained mouse brain coronal sections among 4 genotypes of 6m Q140/Foxp1 KO-het cohort (Bregma 0.65mm and 0.10mm). (I) Quantification of relative ventricle areas in 6m Q140/Foxp1 KO-het cohort (n=3 per genotype).

Article Snippet: PHP1 antibody (Millipore MABN2490; 1:2,000 dilution) and EM48 (Millipore MAB5374; 1:350 dilution) was used for mHTT aggregate detection; Darpp32 antibody (Cell Signaling 2306; 1:500 dilution) and Foxp1 antibody (Abcam ab16645; 1:500 dilution) were used for detecting these targets.

Techniques: Transformation Assay, Expressing, Staining

Journal: bioRxiv

Article Title: Large-scale Perturbation of Systems Biology-Derived Genes Reveals Modifiers of HD- associated Transcriptomic Networks and Pathology

doi: 10.64898/2026.03.02.709091

Figure Lengend Snippet: (A), (B) Z statistics reversal scatterplots for Q140/Scn4b KO-het and Q140/Kcnh4 KO-het. In each plot, x- and y-axes show Z for Q140 vs. WT in our analysis of all Q140 and WT samples and Z for Q140 vs. Q140/Scn4b KO-het (A) and Q140 vs. Q140/Kcnh4 KO-het (B), respectively. Grey lines indicate approximate locations of FDR=0.1 threshold. Blue and red color indicate genes reversed and exacerbated, respectively, at FDR<0.1. Insets indicate numbers of genes in corner regions (DE at FDR<0.1 in both tests). Transcriptome-wide correlation R is shown in a separate inset. (B) Transcriptome-wide Z statistics reversal correlation (Methods) for Q lengths other than Q140 in the allelic series considered a perturbation of Q140 and four other perturbations, namely Q140/Kcnh4 KO-het,, Q140/Pdp1 KO-het, Q140/Bcl11b KO-het and Q140/Scn4b KO-het. (C) Representative images of EM48-stained mHtt aggregates in striata of 6m Q140 and Scn4b/Q140. Coronal striatal sections (30µm thickness) were double stained with anti-Darpp32 (amber) and anti- mHtt (EM48, green) antibodies. (D) Average sizes of nuclear inclusion (NI) in 6m Q140 and Q140/Scn4b KO-het striata (n= 4 per genotype). For all bar graphs in this figure, data are represented as mean ± SEM. Student’s t-test, *** p<0.001. (E) Darpp32 intensities in 6m Q140 and Q140/Scn4b KO-het striata (n= 4 per genotype). Student’s t- test, *p<0.05. (G and H) Representative images (G) and quantification (H) of cells with HTT inclusion bodies (IBs) from four independent HD-MSNs (n = 4) transduced with shControl, shKCNH4, or shSCN4B. Cells were immunostained with anti-mHTT (MW8) and TUBB3 antibodies. An average of 277 cells per sample was counted in three or more randomly chosen fields. Scale bars represent 20 μm (top) and 10 μm (bottom). (I and J) Representative images (I) and quantification (J) of SYTOX-positive cells as a fraction of Hoechst-positive cells in HD-MSNs derived from four independent HD individuals (n = 4) transduced with shControl, shKCNH4, or shSCN4B. Scale bars represent 100 μM. Statistical significance was determined by one-way ANOVA with Tukey’s post hoc test. **p<0.01, *p<0.05, ns, not significant. The sample size (n) corresponds to the number of biologically independent samples; GM04198 (CAG repeat size 47; HD.47), ND33947 (HD.40), ND30013 (HD.43), and GM04230 (HD.45). Each dot represents one individual’s reprogrammed HD-MSN.

Article Snippet: PHP1 antibody (Millipore MABN2490; 1:2,000 dilution) and EM48 (Millipore MAB5374; 1:350 dilution) was used for mHTT aggregate detection; Darpp32 antibody (Cell Signaling 2306; 1:500 dilution) and Foxp1 antibody (Abcam ab16645; 1:500 dilution) were used for detecting these targets.

Techniques: Staining, Transduction, Derivative Assay

Journal: bioRxiv

Article Title: Large-scale Perturbation of Systems Biology-Derived Genes Reveals Modifiers of HD- associated Transcriptomic Networks and Pathology

doi: 10.64898/2026.03.02.709091

Figure Lengend Snippet: (A) Boxplot of log-transformed normalized expression of Pdp1 in the 4 genotypes. Blue points show individual expression values, boxes show the median, median confidence interval, and interquartile range. (B) Reversal scatterplot for Q140/Pdp1 KO-het. X-axis shows Z for Q140 vs. WT in our analysis of all Q140 and WT samples, while the y-axis shows Z for Q140 vs. Q140/Pdp1 KO-het. Grey lines indicate approximate locations of FDR=0.1 threshold. Blue and red color indicate genes reversed and exacerbated, respectively, at FDR<0.1. Insets indicate numbers of genes in corner regions (DE at FDR<0.1 in both tests). Transcriptome-wide correlation R is shown in a separate inset. (C) Enrichment of genes DE in Q140/Pdp1 KO-het vs. Q140 in selected terms that include cell type markers and genes DE in studies of HD models. For clarity, the enrichment p-values are limited to 10-50. (D) Representative images of EM48-stained mHtt aggregates in striata of 6m Q140/Pdp1 KO-het cohort. Coronal striatal sections (30µm thickness) were double stained with anti-Darpp32 (amber) and anti-mHtt (EM48, green) antibodies. (E) Percentage of medium spiny neurons (MSNs) with NIs in striata of 6m Q140 and Q140/Pdp1 KO-het (n= 4 per genotype). For both bar graphs in this figure, data are represented as mean ± SEM. Student’s t- test, **** p<0.0001. (F) Darpp32 intensity in striata of 6m Q140/Pdp1 KO-het cohort (n= 4 per genotype). One-way ANOVA with Tukey’s HSD tests: **** p<0.0001. (G) The top genes and their implicated biological pathways revealed by this large-scale perturbation of mHtt CAG-dependent module hub genes are illustrated here. MSN-selective transcriptionopathy is downstream of somatic mHtt CAG repeat expansion driven by MMR genes (particularly Msh3 and Pms1, ). Genes dysregulated by mHtt – some are selectively expressed in striatal MSNs and others are more broadly expressed - can further exert impacts on MSN-selective pathogenesis in HD mice.

Article Snippet: PHP1 antibody (Millipore MABN2490; 1:2,000 dilution) and EM48 (Millipore MAB5374; 1:350 dilution) was used for mHTT aggregate detection; Darpp32 antibody (Cell Signaling 2306; 1:500 dilution) and Foxp1 antibody (Abcam ab16645; 1:500 dilution) were used for detecting these targets.

Techniques: Transformation Assay, Expressing, Staining