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Figure 7. The pulmonary vascular remodeling and differentiation of lung c-Kit+ cells after Nr2f2 upregulation by AAV (A) Sketch showing the strategy for AAV-Nr2f2 infection of Kit-CreER;Rosa26-tdTomato mice exposed to SuHx. (B) <t>Western</t> <t>blot</t> <t>analysis</t> of Nr2f2 expression in lung c-Kit+ cells after AAV infection. (C) Representative images showing EGFP expression in Kit-CreER;Rosa26-tdTomato mouse lung after AAV-Nr2f2 infection. Br, bronchiole. (D) The RVSP of AAV-infected mice in control and SuHx groups. (E) The RVHI of AAV-infected mice in control and SuHx groups. (F) Representative images showing vascular media of small pulmonary vessels by a-SMA immunostaining (left). Quantification of vascular media thickness (right). (G) Quantification of microvascular muscularization in AAV-infected mice. N, non-muscularization; P, partial muscularization; F, full muscularization. (H) Representative images showing EphB4-positive tdTomato+ ECs in lungs of AAV-infected Kit-CreER;Rosa26-tdTomato mice, with magnification of the boxed region. The circled region in the magnification images shows pulmonary microvessels. Arrows indicate EphB4, tdTomato, and CD31-positive staining cells. Br, bronchiole.
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Figure 7. The pulmonary vascular remodeling and differentiation of lung c-Kit+ cells after Nr2f2 upregulation by AAV (A) Sketch showing the strategy for AAV-Nr2f2 infection of Kit-CreER;Rosa26-tdTomato mice exposed to SuHx. (B) <t>Western</t> <t>blot</t> <t>analysis</t> of Nr2f2 expression in lung c-Kit+ cells after AAV infection. (C) Representative images showing EGFP expression in Kit-CreER;Rosa26-tdTomato mouse lung after AAV-Nr2f2 infection. Br, bronchiole. (D) The RVSP of AAV-infected mice in control and SuHx groups. (E) The RVHI of AAV-infected mice in control and SuHx groups. (F) Representative images showing vascular media of small pulmonary vessels by a-SMA immunostaining (left). Quantification of vascular media thickness (right). (G) Quantification of microvascular muscularization in AAV-infected mice. N, non-muscularization; P, partial muscularization; F, full muscularization. (H) Representative images showing EphB4-positive tdTomato+ ECs in lungs of AAV-infected Kit-CreER;Rosa26-tdTomato mice, with magnification of the boxed region. The circled region in the magnification images shows pulmonary microvessels. Arrows indicate EphB4, tdTomato, and CD31-positive staining cells. Br, bronchiole.
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Figure 7. The pulmonary vascular remodeling and differentiation of lung c-Kit+ cells after Nr2f2 upregulation by AAV (A) Sketch showing the strategy for AAV-Nr2f2 infection of Kit-CreER;Rosa26-tdTomato mice exposed to SuHx. (B) <t>Western</t> <t>blot</t> <t>analysis</t> of Nr2f2 expression in lung c-Kit+ cells after AAV infection. (C) Representative images showing EGFP expression in Kit-CreER;Rosa26-tdTomato mouse lung after AAV-Nr2f2 infection. Br, bronchiole. (D) The RVSP of AAV-infected mice in control and SuHx groups. (E) The RVHI of AAV-infected mice in control and SuHx groups. (F) Representative images showing vascular media of small pulmonary vessels by a-SMA immunostaining (left). Quantification of vascular media thickness (right). (G) Quantification of microvascular muscularization in AAV-infected mice. N, non-muscularization; P, partial muscularization; F, full muscularization. (H) Representative images showing EphB4-positive tdTomato+ ECs in lungs of AAV-infected Kit-CreER;Rosa26-tdTomato mice, with magnification of the boxed region. The circled region in the magnification images shows pulmonary microvessels. Arrows indicate EphB4, tdTomato, and CD31-positive staining cells. Br, bronchiole.
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Figure 7. The pulmonary vascular remodeling and differentiation of lung c-Kit+ cells after Nr2f2 upregulation by AAV (A) Sketch showing the strategy for AAV-Nr2f2 infection of Kit-CreER;Rosa26-tdTomato mice exposed to SuHx. (B) Western blot analysis of Nr2f2 expression in lung c-Kit+ cells after AAV infection. (C) Representative images showing EGFP expression in Kit-CreER;Rosa26-tdTomato mouse lung after AAV-Nr2f2 infection. Br, bronchiole. (D) The RVSP of AAV-infected mice in control and SuHx groups. (E) The RVHI of AAV-infected mice in control and SuHx groups. (F) Representative images showing vascular media of small pulmonary vessels by a-SMA immunostaining (left). Quantification of vascular media thickness (right). (G) Quantification of microvascular muscularization in AAV-infected mice. N, non-muscularization; P, partial muscularization; F, full muscularization. (H) Representative images showing EphB4-positive tdTomato+ ECs in lungs of AAV-infected Kit-CreER;Rosa26-tdTomato mice, with magnification of the boxed region. The circled region in the magnification images shows pulmonary microvessels. Arrows indicate EphB4, tdTomato, and CD31-positive staining cells. Br, bronchiole.

Journal: Developmental cell

Article Title: Differentiation of lung tissue-resident c-Kit + cells into microvascular endothelial cells alleviates pulmonary vascular remodeling.

doi: 10.1016/j.devcel.2025.01.010

Figure Lengend Snippet: Figure 7. The pulmonary vascular remodeling and differentiation of lung c-Kit+ cells after Nr2f2 upregulation by AAV (A) Sketch showing the strategy for AAV-Nr2f2 infection of Kit-CreER;Rosa26-tdTomato mice exposed to SuHx. (B) Western blot analysis of Nr2f2 expression in lung c-Kit+ cells after AAV infection. (C) Representative images showing EGFP expression in Kit-CreER;Rosa26-tdTomato mouse lung after AAV-Nr2f2 infection. Br, bronchiole. (D) The RVSP of AAV-infected mice in control and SuHx groups. (E) The RVHI of AAV-infected mice in control and SuHx groups. (F) Representative images showing vascular media of small pulmonary vessels by a-SMA immunostaining (left). Quantification of vascular media thickness (right). (G) Quantification of microvascular muscularization in AAV-infected mice. N, non-muscularization; P, partial muscularization; F, full muscularization. (H) Representative images showing EphB4-positive tdTomato+ ECs in lungs of AAV-infected Kit-CreER;Rosa26-tdTomato mice, with magnification of the boxed region. The circled region in the magnification images shows pulmonary microvessels. Arrows indicate EphB4, tdTomato, and CD31-positive staining cells. Br, bronchiole.

Article Snippet: Primary antibodies used in western blot analysis were as listed: anti-c-Kit (3074S, CST, 1:1000), anti-Nr2f2 (6434S, CST, 1:1000), anti-DTR (ab66792, Abcam, 1:1000), anti-Scf (ab64677, abcam, 1:1000), anti-b-Actin (AC026, Abclonal, 1:5000).

Techniques: Infection, Western Blot, Expressing, Control, Immunostaining, Staining