Mouse lungs were inflated and fixed in 2% paraformaldehyde, dehydrated in a series of increasing ethanol concentration washes, embedded in paraffin and sectioned. Antibodies used were anti-SM22α (goat anti-SM22α 1:200 Abcam), GFP (goat anti-GFP 1:100 Abcam, rabbit anti-GFP 1:100 Molecular Probe), Scgb1a1 (goat anti-Scgb1a1 1:20 Santa Cruz), SPC (rabbit-anti SPC 1:500 Chemicon), Pdgfrα (rabbit anti-Pdgfrα 1:50 Cell Signaling), Pdgfrβ (rabbit anti-Pdgfrβ 1:100 Cell Signaling), vimentin (rabbit anti-vimentin 1:100 Santa Crux), collagen type1 (rabbit anti-Col1 1:500 Abcam), Ki67 (rabbit anti-Ki67 1:50 Abcam), PCNA (mouse anti-PCNA 1:50 Biocare), PO4-Histone H3 (mouse anti-PO4-Histone H3 1:200 Cell Signaling), TubbIV (mouse anti-TubbIV 1:20 Biogenex), S100A4 (rabbit anti-S100A4 1:200 Abcam). LacZ staining of lungs was performed as previously described8 (link). The slide was imaged on a Zeiss LSM 710 confocal microscope and analyzed in ImageJ software.
Protocol detail
Immunohistochemical Analysis of Mouse Lungs
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anti-sm22α
SM22α
rabbit anti-GFP
Scgb1a1
Pdgfrα
Pdgfrβ
collagen type1
PCNA
mouse anti-PCNA
PO4-Histone H3
mouse anti-PO4-Histone H3
TubbIV
LSM 710 confocal microscope
Antibodies
Collagen
Confocal microscope
Dehydrated ethanol
Embedded paraffin
Goat
Histone h3
Lacz
Lungs
Molecular probe
Mouse
Paraformaldehyde
Pcna
Pdgfrα
Pdgfrβ
Rabbit
Vimentin
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Variable analysis
independent variables
- Mouse lungs were inflated and fixed in 2% paraformaldehyde
- Mouse lungs were dehydrated in a series of increasing ethanol concentration washes
- Mouse lungs were embedded in paraffin and sectioned
- Expression of sm22α (measured by anti-sm22α antibody)
- Expression of GFP (measured by anti-GFP antibodies)
- Expression of Scgb1a1 (measured by anti-Scgb1a1 antibody)
- Expression of SPC (measured by anti-SPC antibody)
- Expression of Pdgfrα (measured by anti-Pdgfrα antibody)
- Expression of Pdgfrβ (measured by anti-Pdgfrβ antibody)
- Expression of vimentin (measured by anti-vimentin antibody)
- Expression of collagen type 1 (measured by anti-Col1 antibody)
- Expression of Ki67 (measured by anti-Ki67 antibody)
- Expression of PCNA (measured by anti-PCNA antibody)
- Expression of PO4-Histone H3 (measured by anti-PO4-Histone H3 antibody)
- Expression of TubbIV (measured by anti-TubbIV antibody)
- Expression of S100A4 (measured by anti-S100A4 antibody)
- LacZ staining of lungs was performed as previously described
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