php eb aav encoding gfp alone  (Addgene inc)

Journal: bioRxiv

Article Title: Soluble DLK1 secreted by telomere-shortening-induced senescent microglia impairs oligodendrocyte functions and alters neuronal activity

doi: 10.64898/2026.01.14.699608

Figure Lengend Snippet: A. Schematic showing the IV injection of the AAV-EGFP or AAV-sDLK1-T2A-GFP into the mice. B. Quantification of the protein levels of sDLK1 in the cortex of the mice, measured by mouse DLK1 ELISA. C. Distributions of the normalized numbers of genes up-regulated (top) and down-regulated (bottom) in different cell types of the mice injected with AAV-sDLK1-T2A-GFP, compared to mice injected with AAV-EGFP. The gene burden score is defined as the number of differentially expressed genes per 1000 UMI detected in each cell type. D. Scatter plot showing the positively correlated genes between DEGs in G3 Terc-/- and AAV-sDLK1-T2A-GFP injected oligodendrocytes. E. Western blot of MBP (top) and MOBP (bottom). F–G. Quantification of the MBP (F) and MOBP (G) western blot. Data were analyzed by two-tailed unpaired t -test. ** p =0.0095 (MBP); *** p =0.0009 (MOBP). H. Ridge plot of the predicted chronological ages for oligodendrocytes in the mice injected with AAV-EGFP (top) and AAV-sDLK1-T2A-GFP (bottom). I. Quantification of the predicted age of oligodendrocytes. Data were reported as a box & whisker plot showing min to max and analyzed by t-test. Each dot represents a cell. **** p < 0.0001. J. UMAP plot showing the enrichment of OPC2 caused by increased sDLK1. K. Ratios of each OPC cluster. Data are reported as mean ± s.e.m. and analyzed by two-way ANOVA. * p =0.0432. L. Running enrichment score and pre-ranked list showing a negative enrichment of oligodendrocyte differentiation predicted by OPC3 markers

Article Snippet: Intravenous injections of AAV particles encoding mouse sDLK1 were performed in C57BL/6 mice, while PHP.eB AAV encoding GFP alone (Addgene, 37825-PHP.eB) served as a negative control.

Techniques: IV Injection, Enzyme-linked Immunosorbent Assay, Injection, Western Blot, Two Tailed Test, Whisker Assay

Journal: bioRxiv

Article Title: Soluble DLK1 secreted by telomere-shortening-induced senescent microglia impairs oligodendrocyte functions and alters neuronal activity

doi: 10.64898/2026.01.14.699608

Figure Lengend Snippet: A. UMAP plots showing eight major cell types identified in the mouse hippocampus. B. Dot plot showing expression levels of canonical cell markers in each identified cell type. C. Proportion of each cell type within animals injected with AAV-GFP and AAV-sDLK1-T2A-GFP. D. Proportion of each cell type within individual samples. E. Violin plots showing the number of unique features (left); the number of total RNA count (middle), and the percentage of mitochondrial genes (right) detected in each identified cell type. F. Correlation between UMI counts and percentage of mitochondrial genes (left) or total gene counts (right) per nuclei for each individual sample.

Article Snippet: Intravenous injections of AAV particles encoding mouse sDLK1 were performed in C57BL/6 mice, while PHP.eB AAV encoding GFP alone (Addgene, 37825-PHP.eB) served as a negative control.

Techniques: Expressing, Injection

Journal: bioRxiv

Article Title: Soluble DLK1 secreted by telomere-shortening-induced senescent microglia impairs oligodendrocyte functions and alters neuronal activity

doi: 10.64898/2026.01.14.699608

Figure Lengend Snippet: A. Dot plot showing the expression level of DLK1 in different cell types in the hippocampus tissue. The size of each dot represents the percentage of cells with detected DLK1 mRNA. B. Chord diagram showing DLK signaling predicted by CellChat. The lengths of the segmented outer circle reflect the expression levels of ligand proteins in each cell type and of receptor proteins in the receiving cells, showing strong expression of DLK1 signaling originating from interneurons and astrocytes to oligodendrocytes and OPCs. C. Bubble plot showing the DLK1 interactions originating from interneurons and astrocytes to different receptors. D. Venn diagram showing the overlap of upregulated DEGs in G3 Terc-/- and AAV-sDLK1-T2A-GFP injected oligodendrocyte(top) and the overlap of downregulated DEGs in G3 Terc-/-and AAV-sDLK1-T2A-GFP injected oligodendrocyte (bottom). E. Dot plot showing the change of expression levels of myelination proteins in oligodendrocytes caused by the increase of sDLK1. F. Cnet plot showing the network of genes associated with myelination-related Gene Ontology terms and myelin-related diseases, based on enrichment analysis of the top 500 differentially expressed genes in oligodendrocytes from mice with AAV-sDLK1-T2A-GFP versus mice with AAV-GFP. Nodes represent genes or GO terms; edge colors represent the pathways each node is involved in.

Article Snippet: Intravenous injections of AAV particles encoding mouse sDLK1 were performed in C57BL/6 mice, while PHP.eB AAV encoding GFP alone (Addgene, 37825-PHP.eB) served as a negative control.

Techniques: Expressing, Injection

Journal: bioRxiv

Article Title: Soluble DLK1 secreted by telomere-shortening-induced senescent microglia impairs oligodendrocyte functions and alters neuronal activity

doi: 10.64898/2026.01.14.699608

Figure Lengend Snippet: A. Representative 20X images of Olig2 in the CA1 region of the hippocampus in C57BL/6 mice injected with either AAV-GFP or AAV-sDLK1-T2A-GFP. Scale bar represents 100 µm. B. Quantification of Olig2+ cell density in CA1. N = 5 mice injected with AAV-GFP and N = 4 mice injected with AAV-sDLK1-T2A-GFP. 2-3 hippocampal sections/mouse were imaged and analyzed. Data are reported as mean ± SEM. p = 0.9480. Data were analyzed by unpaired t-test. C. Quantification of Olig2+ cell density in CA3. N = 5 mice injected with AAV-GFP and N = 4 mice injected with AAV-sDLK1-T2A-GFP. 2-3 hippocampal sections/mouse were imaged and analyzed. Data are reported as mean ± SEM. p = 0.2008. Data were analyzed by unpaired t-test. D. Representative 20X images of PDGFRα in the CA1 region of the hippocampus in C57BL/6 mice injected with either AAV-GFP or AAV-sDLK1-T2A-GFP. Scale bar represents 100 µm. E. Quantification of PDGFRα+ cell density in CA1. N = 5 mice/treatment group and 2-3 hippocampal sections/mouse were imaged and analyzed. Data are reported as mean ± SEM. p = 0.3534. Data were analyzed by unpaired t-test. F. Quantification of PDGFRα+ cell density in CA3. N = 5 mice/treatment group and 2-3 hippocampal sections/mouse were imaged and analyzed. Data are reported as mean ± SEM. p = 0.6744. Data were analyzed by unpaired t-test.

Article Snippet: Intravenous injections of AAV particles encoding mouse sDLK1 were performed in C57BL/6 mice, while PHP.eB AAV encoding GFP alone (Addgene, 37825-PHP.eB) served as a negative control.

Techniques: Injection

Journal: bioRxiv

Article Title: Soluble DLK1 secreted by telomere-shortening-induced senescent microglia impairs oligodendrocyte functions and alters neuronal activity

doi: 10.64898/2026.01.14.699608

Figure Lengend Snippet: A Venn diagram showing the overlap of upregulated (top) and downregulated (bottom) DEGs in G3 Terc-/- and AAV-sDLK1-T2A-GFP -injected excitatory neurons. B. Dot plot of the top 10 Reactome pathways inferred by the upregulated overlapping DEGs in G3 Terc-/- and AAV-sDLK1-T2A-GFP injected excitatory neurons. C–D. Running enrichment score and pre-ranked list showing a positive (C) and negative (D) enrichment of calcium ion transmembrane transport predicted by the upregulated overlapping DEGs in G3 Terc-/- and AAV-sDLK1-T2A-GFP injected excitatory neurons. E. Schematic illustrating the experiment setup to test the chronic effects of DLK1 on neuronal activities. F. Representative fluorescence image of human iPSC-derived neurons expressing GCaMP8f showing spontaneous activity. G. Representative spontaneous calcium traces. H–J. Quantification of synchronized firing rate (H). firing amplitude (I), or spontaneous firing rate (J). K. Representative averaged calcium traces from one KCl stimulation experiment in neurons treated with DLK1 (red) and the untreated control neurons (black). Recording 400 seconds. Mean± s.e.m. L–N. Quantification of peak amplitude (L), the delayed KCl stimulation-induced neuronal responses (M), the time each neuron spent to reach peak intensity (N), from KCl stimulation-induced neuronal responses. Data are presented as mean ± s.e.m. and analyzed by unpaired t -test. ** p =0.0055 (L) **** p <0.0001 (N).

Article Snippet: Intravenous injections of AAV particles encoding mouse sDLK1 were performed in C57BL/6 mice, while PHP.eB AAV encoding GFP alone (Addgene, 37825-PHP.eB) served as a negative control.

Techniques: Injection, Fluorescence, Derivative Assay, Expressing, Activity Assay, Control