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a, Schematic diagram illustrating major airway cell types, their anatomical locations, and antibody markers. Mouse tissues were collected 21 days after intranasal administration of 1 × 10 11 vg of AAV-CAG-GFP. b, Representative images showing transduced goblet cells <t>(MUC5A</t> + ) in the nasal cavity. Scale bar: 25 μm for low-magnification images and 5 μm for enlarged images. c, AAV-mediated transduction of club cells (CC10 + ) in the trachea. Arrows point to cells co-labeled with GFP and CC10 antibodies. Scale bar: 25 μm. d , AAV-mediated transduction of alveolar type I (AT1) (RAGE + ) in lung parenchyma tissues. Scale bar: 25 μm. e-g , Quantifications on the percentage of GFP + goblet cells in the nasal cavity (e), club cells in the trachea (f) and AT1 cells in the lung (g). Mean ± s.e.m., n = 3-4 for each group. Statistical analyses were performed using one-way ANOVA with Tukey’s multiple comparison test (** p < 0.01, *** p < 0.001).
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a, Schematic diagram illustrating major airway cell types, their anatomical locations, and antibody markers. Mouse tissues were collected 21 days after intranasal administration of 1 × 10 11 vg of AAV-CAG-GFP. b, Representative images showing transduced goblet cells <t>(MUC5A</t> + ) in the nasal cavity. Scale bar: 25 μm for low-magnification images and 5 μm for enlarged images. c, AAV-mediated transduction of club cells (CC10 + ) in the trachea. Arrows point to cells co-labeled with GFP and CC10 antibodies. Scale bar: 25 μm. d , AAV-mediated transduction of alveolar type I (AT1) (RAGE + ) in lung parenchyma tissues. Scale bar: 25 μm. e-g , Quantifications on the percentage of GFP + goblet cells in the nasal cavity (e), club cells in the trachea (f) and AT1 cells in the lung (g). Mean ± s.e.m., n = 3-4 for each group. Statistical analyses were performed using one-way ANOVA with Tukey’s multiple comparison test (** p < 0.01, *** p < 0.001).
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a, Schematic diagram illustrating major airway cell types, their anatomical locations, and antibody markers. Mouse tissues were collected 21 days after intranasal administration of 1 × 10 11 vg of AAV-CAG-GFP. b, Representative images showing transduced goblet cells <t>(MUC5A</t> + ) in the nasal cavity. Scale bar: 25 μm for low-magnification images and 5 μm for enlarged images. c, AAV-mediated transduction of club cells (CC10 + ) in the trachea. Arrows point to cells co-labeled with GFP and CC10 antibodies. Scale bar: 25 μm. d , AAV-mediated transduction of alveolar type I (AT1) (RAGE + ) in lung parenchyma tissues. Scale bar: 25 μm. e-g , Quantifications on the percentage of GFP + goblet cells in the nasal cavity (e), club cells in the trachea (f) and AT1 cells in the lung (g). Mean ± s.e.m., n = 3-4 for each group. Statistical analyses were performed using one-way ANOVA with Tukey’s multiple comparison test (** p < 0.01, *** p < 0.001).
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a, Schematic diagram illustrating major airway cell types, their anatomical locations, and antibody markers. Mouse tissues were collected 21 days after intranasal administration of 1 × 10 11 vg of AAV-CAG-GFP. b, Representative images showing transduced goblet cells <t>(MUC5A</t> + ) in the nasal cavity. Scale bar: 25 μm for low-magnification images and 5 μm for enlarged images. c, AAV-mediated transduction of club cells (CC10 + ) in the trachea. Arrows point to cells co-labeled with GFP and CC10 antibodies. Scale bar: 25 μm. d , AAV-mediated transduction of alveolar type I (AT1) (RAGE + ) in lung parenchyma tissues. Scale bar: 25 μm. e-g , Quantifications on the percentage of GFP + goblet cells in the nasal cavity (e), club cells in the trachea (f) and AT1 cells in the lung (g). Mean ± s.e.m., n = 3-4 for each group. Statistical analyses were performed using one-way ANOVA with Tukey’s multiple comparison test (** p < 0.01, *** p < 0.001).
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a, Schematic diagram illustrating major airway cell types, their anatomical locations, and antibody markers. Mouse tissues were collected 21 days after intranasal administration of 1 × 10 11 vg of AAV-CAG-GFP. b, Representative images showing transduced goblet cells (MUC5A + ) in the nasal cavity. Scale bar: 25 μm for low-magnification images and 5 μm for enlarged images. c, AAV-mediated transduction of club cells (CC10 + ) in the trachea. Arrows point to cells co-labeled with GFP and CC10 antibodies. Scale bar: 25 μm. d , AAV-mediated transduction of alveolar type I (AT1) (RAGE + ) in lung parenchyma tissues. Scale bar: 25 μm. e-g , Quantifications on the percentage of GFP + goblet cells in the nasal cavity (e), club cells in the trachea (f) and AT1 cells in the lung (g). Mean ± s.e.m., n = 3-4 for each group. Statistical analyses were performed using one-way ANOVA with Tukey’s multiple comparison test (** p < 0.01, *** p < 0.001).

Journal: bioRxiv

Article Title: Tropism of AAV.CPP.16 in the respiratory tract and its application for a CRISPR-based gene therapy against SARS-CoV-2

doi: 10.1101/2023.11.17.567583

Figure Lengend Snippet: a, Schematic diagram illustrating major airway cell types, their anatomical locations, and antibody markers. Mouse tissues were collected 21 days after intranasal administration of 1 × 10 11 vg of AAV-CAG-GFP. b, Representative images showing transduced goblet cells (MUC5A + ) in the nasal cavity. Scale bar: 25 μm for low-magnification images and 5 μm for enlarged images. c, AAV-mediated transduction of club cells (CC10 + ) in the trachea. Arrows point to cells co-labeled with GFP and CC10 antibodies. Scale bar: 25 μm. d , AAV-mediated transduction of alveolar type I (AT1) (RAGE + ) in lung parenchyma tissues. Scale bar: 25 μm. e-g , Quantifications on the percentage of GFP + goblet cells in the nasal cavity (e), club cells in the trachea (f) and AT1 cells in the lung (g). Mean ± s.e.m., n = 3-4 for each group. Statistical analyses were performed using one-way ANOVA with Tukey’s multiple comparison test (** p < 0.01, *** p < 0.001).

Article Snippet: Primary antibodies included: chicken anti-GFP (1:1000, AB13970, Abcam), mouse anti-GFP (1:3000, GB12602, Servicebio); rabbit anti-RFP (1:1000, 600-401-379, Rockland), rabbit anti-α-Tubulin (1:400, 5335, Cell signaling technology), rabbit anti-α-Tubulin (1:200, D20G3, Cell signaling technology), mouse anti-MUC5A (1:400, MA5-12178, Invitrogen), mouse anti-CC10 (1:400, sc-390313, Santa Cruz), rabbit anti-CC10 (1:200, A16997, abclonal), rat anti-RAGE (1:400, MAB1179-SP, R&D), rabbit anti-RAGE (1:200, 16346-1-AP, Proteintech).

Techniques: Transduction, Labeling, Comparison