Keratinocyte growth factor protects against Clara cell injury induced by naphthalene

Supplementary methods and results

Materials and methods: Kinetics of naphthalene (NA)-induced acute airway injury; RNA isolation and Real-Time Reverse Transcriptase (RT-PCR); Double immunohistochemistry. Results: Kinetics of acute airway injury induced by naphthalene. Figures: Hematoxilin-Eosin staining of distal airways of mice; Double immunohistochemistry for ciliated cells and Clara cells. Table: Primers for quantitative Real-Time Reverse Transcriptase-PCR

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• Supplementary methods and results - Materials and methods: Kinetics of naphthalene (NA)-induced acute airway injury; RNA isolation and Real-Time Reverse Transcriptase (RT-PCR); Double immunohistochemistry. Results: Kinetics of acute airway injury induced by naphthalene. Table: Primers for quantitative Real-Time Reverse Transcriptase-PCR
• Figure E1 - FIGURE E1. Hematoxilin-Eosin staining of distal airways of mice. Mice were treated with naphthalene (200 mg/kg) and sacrificed after 2, 3, 6, 8, 12, 16, and 24 h. Nucleus of cells are stained dark-blue. Clara cells (arrow) as well as ciliated cells (black asterisk) are visible in the corn oil treated mice (Fig.E1 A). Mice, which were treated for 2 h (Fig.E1 B) with naphthalene (NA), reveal no morphological alteration. Slight vacuolation (white asterisk) and swelling (arrow) is seen at 3 h (Fig.E1 C) and 6 h (Fig.E1 D) after NA-injection. At 8 h (Fig.E1 E) Clara cells begin to exfoliate (arrow) and severity of morphological alterations increased at 12 h (Fig.E1 F) and 16 h (Fig.E1 F). At 24 h (Fig.E1 H) most of the Clara cells are lost by exfoliation and the basal lamina is covered by a layer of squamous epithelial cells. Scale bar 12 �m.
• Figure E2 - FIGURE E2. Double immunohistochemistry for ciliated cells and Clara cells. The distal airway epithelium of mice was almost exclusively composed of Clara cells (green) and ciliated cells (brown). Sections were counterstained with hematoxylin. Scale bar 100 �m.