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ITGA3 regulates AT2 cell function via FAK/YAP signaling in response to CSE . ( A-D ) Western blot analysis of FAK, p-FAK, ITGA3, n-YAP of Itga3-overexpressed and NC-infected organoid with or without 2.5 % CSE treatment. β-actin and Lamin B1 was separately used as loading control of total protein and nuclear protein. ( E ) Immunofluorescence staining of n-YAP (white), pro-SPC (red) and DAPI (blue) of Itga3-overexpressed and NC-infected organoid with or without 2.5 % CSE treatment. Scale bar = 100 μm. ( F ) Quantification of n-YAP immunofluorescence intensity. ( G ) Representative bright field (scale bar = 500 μm) and immunostaining images (scale bar = 100 μm) of NC-infected and Itga3-overexpressed organoid with 2.5 % CSE treatment, and Itga3-overexpressed organoid with 2.5 % CSE and 2 μM Y15, 1 μM VP, separately. Ki67 (white), pro-SPC (red) and DAPI (blue). (H-I ) Quantification of forming efficiency and diameter of organoid in (G). ( J ) Percentage of Ki67 + pro-SPC + cells among all pro-SPC + cells in (G). Data are represented as mean ± SD. Statistical significance *P < 0.05, **P < 0.01, ***P < 0.001. p-FAK, phospho-FAK. n-YAP, nuclear YAP. VP, <t>Verteporfin.</t>
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ITGA3 regulates AT2 cell function via FAK/YAP signaling in response to CSE . ( A-D ) Western blot analysis of FAK, p-FAK, ITGA3, n-YAP of Itga3-overexpressed and NC-infected organoid with or without 2.5 % CSE treatment. β-actin and Lamin B1 was separately used as loading control of total protein and nuclear protein. ( E ) Immunofluorescence staining of n-YAP (white), pro-SPC (red) and DAPI (blue) of Itga3-overexpressed and NC-infected organoid with or without 2.5 % CSE treatment. Scale bar = 100 μm. ( F ) Quantification of n-YAP immunofluorescence intensity. ( G ) Representative bright field (scale bar = 500 μm) and immunostaining images (scale bar = 100 μm) of NC-infected and Itga3-overexpressed organoid with 2.5 % CSE treatment, and Itga3-overexpressed organoid with 2.5 % CSE and 2 μM Y15, 1 μM VP, separately. Ki67 (white), pro-SPC (red) and DAPI (blue). (H-I ) Quantification of forming efficiency and diameter of organoid in (G). ( J ) Percentage of Ki67 + pro-SPC + cells among all pro-SPC + cells in (G). Data are represented as mean ± SD. Statistical significance *P < 0.05, **P < 0.01, ***P < 0.001. p-FAK, phospho-FAK. n-YAP, nuclear YAP. VP, <t>Verteporfin.</t>
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ITGA3 regulates AT2 cell function via FAK/YAP signaling in response to CSE . ( A-D ) Western blot analysis of FAK, p-FAK, ITGA3, n-YAP of Itga3-overexpressed and NC-infected organoid with or without 2.5 % CSE treatment. β-actin and Lamin B1 was separately used as loading control of total protein and nuclear protein. ( E ) Immunofluorescence staining of n-YAP (white), pro-SPC (red) and DAPI (blue) of Itga3-overexpressed and NC-infected organoid with or without 2.5 % CSE treatment. Scale bar = 100 μm. ( F ) Quantification of n-YAP immunofluorescence intensity. ( G ) Representative bright field (scale bar = 500 μm) and immunostaining images (scale bar = 100 μm) of NC-infected and Itga3-overexpressed organoid with 2.5 % CSE treatment, and Itga3-overexpressed organoid with 2.5 % CSE and 2 μM Y15, 1 μM VP, separately. Ki67 (white), pro-SPC (red) and DAPI (blue). (H-I ) Quantification of forming efficiency and diameter of organoid in (G). ( J ) Percentage of Ki67 + pro-SPC + cells among all pro-SPC + cells in (G). Data are represented as mean ± SD. Statistical significance *P < 0.05, **P < 0.01, ***P < 0.001. p-FAK, phospho-FAK. n-YAP, nuclear YAP. VP, <t>Verteporfin.</t>
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ITGA3 regulates AT2 cell function via FAK/YAP signaling in response to CSE . ( A-D ) Western blot analysis of FAK, p-FAK, ITGA3, n-YAP of Itga3-overexpressed and NC-infected organoid with or without 2.5 % CSE treatment. β-actin and Lamin B1 was separately used as loading control of total protein and nuclear protein. ( E ) Immunofluorescence staining of n-YAP (white), pro-SPC (red) and DAPI (blue) of Itga3-overexpressed and NC-infected organoid with or without 2.5 % CSE treatment. Scale bar = 100 μm. ( F ) Quantification of n-YAP immunofluorescence intensity. ( G ) Representative bright field (scale bar = 500 μm) and immunostaining images (scale bar = 100 μm) of NC-infected and Itga3-overexpressed organoid with 2.5 % CSE treatment, and Itga3-overexpressed organoid with 2.5 % CSE and 2 μM Y15, 1 μM VP, separately. Ki67 (white), pro-SPC (red) and DAPI (blue). (H-I ) Quantification of forming efficiency and diameter of organoid in (G). ( J ) Percentage of Ki67 + pro-SPC + cells among all pro-SPC + cells in (G). Data are represented as mean ± SD. Statistical significance *P < 0.05, **P < 0.01, ***P < 0.001. p-FAK, phospho-FAK. n-YAP, nuclear YAP. VP, <t>Verteporfin.</t>
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ITGA3 regulates AT2 cell function via FAK/YAP signaling in response to CSE . ( A-D ) Western blot analysis of FAK, p-FAK, ITGA3, n-YAP of Itga3-overexpressed and NC-infected organoid with or without 2.5 % CSE treatment. β-actin and Lamin B1 was separately used as loading control of total protein and nuclear protein. ( E ) Immunofluorescence staining of n-YAP (white), pro-SPC (red) and DAPI (blue) of Itga3-overexpressed and NC-infected organoid with or without 2.5 % CSE treatment. Scale bar = 100 μm. ( F ) Quantification of n-YAP immunofluorescence intensity. ( G ) Representative bright field (scale bar = 500 μm) and immunostaining images (scale bar = 100 μm) of NC-infected and Itga3-overexpressed organoid with 2.5 % CSE treatment, and Itga3-overexpressed organoid with 2.5 % CSE and 2 μM Y15, 1 μM VP, separately. Ki67 (white), pro-SPC (red) and DAPI (blue). (H-I ) Quantification of forming efficiency and diameter of organoid in (G). ( J ) Percentage of Ki67 + pro-SPC + cells among all pro-SPC + cells in (G). Data are represented as mean ± SD. Statistical significance *P < 0.05, **P < 0.01, ***P < 0.001. p-FAK, phospho-FAK. n-YAP, nuclear YAP. VP, Verteporfin.

Journal: Journal of Advanced Research

Article Title: Dysregulated ITGA3/FAK/YAP axis mediates impaired alveolar type II epithelial cells function in COPD

doi: 10.1016/j.jare.2025.11.033

Figure Lengend Snippet: ITGA3 regulates AT2 cell function via FAK/YAP signaling in response to CSE . ( A-D ) Western blot analysis of FAK, p-FAK, ITGA3, n-YAP of Itga3-overexpressed and NC-infected organoid with or without 2.5 % CSE treatment. β-actin and Lamin B1 was separately used as loading control of total protein and nuclear protein. ( E ) Immunofluorescence staining of n-YAP (white), pro-SPC (red) and DAPI (blue) of Itga3-overexpressed and NC-infected organoid with or without 2.5 % CSE treatment. Scale bar = 100 μm. ( F ) Quantification of n-YAP immunofluorescence intensity. ( G ) Representative bright field (scale bar = 500 μm) and immunostaining images (scale bar = 100 μm) of NC-infected and Itga3-overexpressed organoid with 2.5 % CSE treatment, and Itga3-overexpressed organoid with 2.5 % CSE and 2 μM Y15, 1 μM VP, separately. Ki67 (white), pro-SPC (red) and DAPI (blue). (H-I ) Quantification of forming efficiency and diameter of organoid in (G). ( J ) Percentage of Ki67 + pro-SPC + cells among all pro-SPC + cells in (G). Data are represented as mean ± SD. Statistical significance *P < 0.05, **P < 0.01, ***P < 0.001. p-FAK, phospho-FAK. n-YAP, nuclear YAP. VP, Verteporfin.

Article Snippet: 2 μM Y15 (HY-12444, MedChemExpress, USA), 1 μM Verteporfin (VP) (HY-B0146, MedChemExpress, USA), 1 μM PY-60 (HY-141644, MedChemExpress, USA), 0.5 mM NAC (HY-B0215, MedChemExpress, USA), were used in organoid pharmacological assay, separately.

Techniques: Cell Function Assay, Western Blot, Infection, Control, Immunofluorescence, Staining, Immunostaining