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. 2007 Mar 15;37(1):67–74. doi: 10.1165/rcmb.2006-0469OC

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Figure 7. — Increased apoptosis in lung sections from HPS mice after intratracheal bleomycin challenge. At indicated time points after intratracheal bleomycin (0.025 U), lungs were inflation-fixed in formalin as previously described. TUNEL assay (Roche) was performed on paraffin-embedded sections. (A–C) Representative fluorescent micrographs (×40) for (A) WT, (B) Pearl, and (C) Pale Ear at 5 h after bleomycin challenge are shown. (D–F) Representative co-immunofluorescence for TUNEL (D) and pro–SP-C (E) identifies apoptotic alveolar type II cells (F, merge) from a Pearl lung section, 5 h after intratracheal bleomycin challenge. (G and H) Representative photomicrographs (×40) of active caspase 3 staining in lung sections from (G) WT, (H) Pearl, or (I) Pale Ear mice, 3 d after intratracheal bleomycin challenge. Increased numbers of cells positive for active caspase 3 were present in the lungs of Pearl and Pale mice. Arrows indicate examples of positive cells. (J) Quantitative assessment of percentage of alveolar type II cells which are TUNEL-positive. Alveolar wall TUNEL-positive and SP-C–positive cells were counted from 10 high-power (×40) fields (n = 3 each at baseline and 5 h, n = 4 each at 24 h, *P < 0.05). (K) Levels of Fas ligand in the lungs of Pearl mice after intratracheal bleomycin challenge. Mice were challenged with intratracheal bleomycin (0.025 U). Mice were killed at various time points, lungs were harvested, and Fas ligand (FasL) levels were assessed by ELISA (n = 4 per group, *P < 0.05 for Pearl versus WT, Day 7).

Figure 7. — Increased apoptosis in lung sections from HPS mice after intratracheal bleomycin challenge. At indicated time points after intratracheal bleomycin (0.025 U), lungs were inflation-fixed in formalin as previously described. TUNEL assay (Roche) was performed on paraffin-embedded sections. (A–C) Representative fluorescent micrographs (×40) for (A) WT, (B) Pearl, and (C) Pale Ear at 5 h after bleomycin challenge are shown. (D–F) Representative co-immunofluorescence for TUNEL (D) and pro–SP-C (E) identifies apoptotic alveolar type II cells (F, merge) from a Pearl lung section, 5 h after intratracheal bleomycin challenge. (G and H) Representative photomicrographs (×40) of active caspase 3 staining in lung sections from (G) WT, (H) Pearl, or (I) Pale Ear mice, 3 d after intratracheal bleomycin challenge. Increased numbers of cells positive for active caspase 3 were present in the lungs of Pearl and Pale mice. Arrows indicate examples of positive cells. (J) Quantitative assessment of percentage of alveolar type II cells which are TUNEL-positive. Alveolar wall TUNEL-positive and SP-C–positive cells were counted from 10 high-power (×40) fields (n = 3 each at baseline and 5 h, n = 4 each at 24 h, *P < 0.05). (K) Levels of Fas ligand in the lungs of Pearl mice after intratracheal bleomycin challenge. Mice were challenged with intratracheal bleomycin (0.025 U). Mice were killed at various time points, lungs were harvested, and Fas ligand (FasL) levels were assessed by ELISA (n = 4 per group, *P < 0.05 for Pearl versus WT, Day 7).