Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2023 Nov 16;14:7426. doi: 10.1038/s41467-023-42746-w

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 5 — a Immunohistochemical analysis of RiboTag⁺ cells (hemagglutinin HA⁺ ribosomal tag), colocalized with astrocytic marker Aldh1l1 and astrocytic reactivity marker GFAP. Astrocytic specificity is high in Aldh1l1-CreERT2 transgenic mice and systemic delivery of PHP.eB::GfaABC1D-Cre-4x6T with no overt evidence of astrocyte reactivity (GFAP% coverage). Specificity remains high with intracortical delivery of PHP.eB::GfaABC1D-Cre-4x6T and decreases with intracortical delivery of AAV2/5::GfaABC1D-Cre; this route of viral delivery shows some evidence of astrocyte reactivity by GFAP immunoreactivity. Scale bars: 50 μm. Mean ± SEM; n = 4 mice per cohort. Astrocyte specificity, HA⁺Aldh1l1⁺/HA⁺: one-way ANOVA, Holm–Sidak’s multiple comparisons test, P < 0.0001, F = 77.01, df=15; Aldh1l1-CreERT2 vs AAV2/5::GfaABC1D-Cre ****P < 0.0001. GFAP % area coverage: Aldh1l1-CreERT2 = 2.33% coverage ± 0.64; PHP.eB::GfaABC1D-Cre-4x6T systemic, 2.77% coverage ± 0.55; PHP.eB::GfaABC1D-Cre-4x6T intracortical, 13.61% coverage ± 4.71; AAV2/5::GfaABC1D-Cre, 12.52% coverage ± 2.90; one-way ANOVA, Holm–Sidak’s multiple comparisons test, P = 0.0011, F = 10.54, df=15; Aldh1l1-CreERT2 vs PHP.eB::GfaABC1D-Cre-4x6T intracortical *P = 0.0431; Aldh1l1-CreERT2 vs AAV2/5::GfaABC1D-Cre *P = 0.0481. Source data are provided as a Source Data file. b Relative levels of enrichment and de-enrichment of canonical genes for astrocytes, neurons, oligodendrocytes, microglia, and endothelial cells in IP-vs-input samples. c Differentially expressed genes in IP samples: different viral cohorts vs Aldh1l1-CreERT2 IP samples, and systemic vs cortical 4x6T samples. FDR < 0.05, average FPKM across all IP samples >1. d Cell-type specificity of differentially expressed genes in IP samples: genes in (c) that are represented in the top 1000 specific genes for astrocytes, endothelial cells, microglia, neurons, or oligodendrocytes. e Venn diagram showing the overlap of which neuron-specific genes from (d) are upregulated in different viral cohort IP samples vs Aldh1l1-CreERT2 samples. f Top ten most significantly upregulated and downregulated Gene Ontology gene sets (FDR < 0.05) in ranked IP transcriptomes of each of the viral cohorts, calculated with Gene Set Enrichment Analysis; some gene sets overlapped, particularly for 4x6T cohorts, and only seven gene sets passed FDR < 0.05 for intracortical 4x6T. In cases where the FDR was 0, scores were reassigned as 0.0001 to visualize the relative -logFDR. g Numbers of genes potentially regulated by miRNAs that comprise the 4x6T cassette (5409 total) that show evidence of 4x6T-specific regulation in IP samples (FDR < 0.05 in either 4x6T cohort vs Aldh1l1-CreERT2 cohort; not differentially regulated in AAV2/5 cohort vs Aldh1l1-CreERT2 cohort). Note: none of the 5409 genes show evidence of 4x6T-specific regulation in input samples. n = 4 mice per cohort; 2–4 months old, euthanized 2 weeks after final tamoxifen administration and 18 days postvirus injection. Titer: retroorbital PHP.eB::GfaABC1D-Cre-4x6T, 1 × 10¹² vg/mouse; intracortical, PHP.eB::GfaABC1D-Cre-4x6T or AAV2/5::GfaABC1D-Cre: 500 nl of 1 × 10¹² vg/ml in each of three sites.