hamster anti t1a  (Developmental Studies Hybridoma Bank)

Journal:

Article Title: Lung alveolar integrity is compromised by telomere shortening in telomerase-null mice

doi: 10.1152/ajplung.90411.2008

Figure Lengend Snippet: Analysis of Clara cell secretory protein (CCSP)-positive airway cells, T1a-positive AEC1, and surfactant protein-C (SP-C)-positive AEC2 in normoxia-maintained WT F4 and terc−/− F4 samples in situ. A: lung tissue from WT F4 and terc−/− F4 mice maintained in normoxia was analyzed for CCSP and SP-C expression by immunohistochemistry. To control for nonspecific antibody staining, purified rabbit or hamster IgG was used at the same concentration as primary anti-CCSP, anti-T1a, and anti-SP-C antibodies (not shown). Sections were fixed and subjected to immunohistochemistry with the primary antibodies indicated and a Cy3-labeled anti-rabbit IgG secondary antibody. Top: CCSP-positive cells are shown lining the large airways in each sample. Middle: long, thin, T1a-positive AEC1 can be observed lining alveolar walls. Bottom: arrows indicate brightly staining SP-C-positive AEC2 scattered through lung parenchyma. In the WT F4 section, arrows point to a portion of all SP-C-positive cells, while in the terc−/− F4 section, arrows point to all SP-C-positive cells identified. All panels were observed at ×20. B: quantitation of SP-C positive AEC2 present in WT F4 and terc−/− F4 lung tissue. SP-C-labeled sections were observed microscopically, and the number of SP-C-positive cells was counted per microscopic field at ×20. For each sample, n = 8. The mean number of SP-C positive cells per field in normoxic WT lung was 42.6 (SE ±2.88), while in normoxic terc−/− F4 lung, the number was 19.2 (SE ±6.53). The 2-tailed P value for comparison of these populations was highly significant (*P = 0.0095) C: quantitation of SP-C-positive AEC2 present in WT F4 and terc−/−F4 lung tissue as % of total cell number. SP-C-labeled sections were observed microscopically, and the number of SP-C-positive cells was counted per total number of cells (identified by positive DAPI staining) per microscopic field at ×20. For each sample, n = 8. Mean % of SP-C positive cells per field in normoxic WT lung was 12.4% (SE ±0.89), while in normoxic terc−/−F4 lung, the mean was 10.6% (SE ±4.39). The 2-tailed P value for comparison of these populations was not significant (P = 0.6773).

Article Snippet: Syrian hamster monoclonal antibody to T1a (hybridoma no. 8.1.1) was from the Developmental Studies Hybridoma Bank, University of Iowa.

Techniques: In Situ, Expressing, Immunohistochemistry, Control, Staining, Purification, Concentration Assay, Labeling, Quantitation Assay, Comparison

Journal:

Article Title: Lung alveolar integrity is compromised by telomere shortening in telomerase-null mice

doi: 10.1152/ajplung.90411.2008

Figure Lengend Snippet: terc−/− F4 lung tissue exhibits markers for DNA damage under normoxic conditions. A: TUNEL in situ of normoxia-maintained WT F4 and terc−/− F4 samples. Lung tissue samples were subjected to TUNEL to detect cells carrying DNA strand breaks. FITC-labeled dUTP incorporated into damaged DNA appears green, while tissue cell nuclei were stained blue with DAPI. Sections were observed at ×20. B: quantitation of TUNEL in situ of normoxia-maintained WT F4 and terc−/− F4 samples. TUNEL-labeled sections were observed microscopically, and the number of TUNEL-positive cells was counted per total number of cells (by DAPI) in each microscopic field at ×20. Fields were counted from sections obtained the left lobe of each lung. For each sample, n = 3. *P = 0.0044 by Student's t-test. C: 8-oxoguanine (8-OHdG, 8-oxo-dG) immunohistochemical analysis of normoxia-maintained WT F4 and terc−/− F4 samples and colocalization with SP-C expression. Top: lung tissue from WT F4 and terc−/− F4 mice maintained in normoxia was analyzed for 8-OHdG expression by immunohistochemistry. Green arrows indicate brightly staining OHdG-positive cells. Middle: immunohistochemistry for 8-OHdG was combined with staining for T1a expression. T1a-positive and 8-OHdG double-positive cells are indicated by green arrows. Bottom: immunohistochemistry for 8-OHdG was combined with staining for SP-C expression. SP-C-positive cells and OHdG-positive cells are indicated by red and green arrows, respectively. Yellow arrows point to red, SP-C-positive cells with cytoplasmic expression (green) of 8-OHdG. Sections were observed at ×40. D: quantitation of 8-OHdG in situ of normoxia-maintained WT F4 and terc−/− F4 samples. 8-OHdG-labeled sections were observed microscopically, and the number of OHdG-positive cells was counted per whole left lobe section. Six sections from 2 individual animals were analyzed for each sample (n = 6). *P = 0.0042 by Student's t-test.

Article Snippet: Syrian hamster monoclonal antibody to T1a (hybridoma no. 8.1.1) was from the Developmental Studies Hybridoma Bank, University of Iowa.

Techniques: TUNEL Assay, In Situ, Labeling, Staining, Quantitation Assay, Immunohistochemical staining, Expressing, Immunohistochemistry