Figure 6.

Inflammatory parameters in Muc5ac^+/+ and Muc5ac^−/− acute lung injury. Muc5ac^−/− mice or their corresponding littermate controls matched in age, sex, and weight were mechanically ventilated (inspiratory pressure (P_insp) 45 mbar, inspired oxygen fraction 1.0, positive-end expiratory pressure 3 mbar). After the indicated time points, the lungs were harvested and snap frozen in liquid nitrogen. (a) Frozen tissue was homogenized and probed for myeloperoxidase (MPO) protein levels by enzyme-linked immunosorbent assay (ELISA). MPO content was normalized to total protein content of homogenate determined by bicinchoninic acid assay (BCA). Data are expressed as mean ± s.d. (n = 5). (b) Total RNA was isolated and Cxcl1 mRNA levels were determined by real-time reverse transcriptase–PCR (RT-PCR). Data were calculated relative to the internal housekeeping gene (β-actin) and are expressed as mean fold change compared with control (0 h ventilation) ± s.d. (n = 4). (c) Frozen tissue was homogenized and probed for Cxcl1 protein levels by ELISA. Cxcl1 content was normalized to total protein content of homogenate determined by BCA. Data are expressed as mean ± s.d. (n= 5). (d) Total RNA was isolated and Cxcl2 mRNA levels were determined by real-time RT-PCR. Data were calculated relative to the internal housekeeping gene (β-actin) and are expressed as mean fold change compared with control (0 h ventilation) ± s.d. (e) Frozen tissue was homogenized and probed for interleukin (IL)-6 protein levels by ELISA. IL-6 content was normalized to total protein content of homogenate determined by BCA. Data are expressed as mean ± s.d. (n = 5).