Impaired Resolution of Inflammation in the Endoglin Heterozygous Mouse Model of Chronic Colitis
Figure 4
Reduced colonic MPO and Nox-2 expression in colitic Eng +/− mice are not associated with an intrinsic bone marrow defect. (a) Representative Western blots show colonic expression levels of MPO and Nox-2 in basal and colitic mice, normalized to β-actin (days 0 and 18–23 images are derived from a single immunoblot). Histograms illustrate densitometric analysis of (b) MPO and (c) Nox-2 expression (for MPO: –8 mice for day 0 and 4–9 mice for days 7–9 and 18–23; for Nox-2: -5 mice for day 0, 3–7 for days 7–9 and 18–23). (d) Colonic MPO enzymatic activity was assessed in Eng +/+ and Eng +/− mice at days 0, 9–12, and 19 of colitis (–8 mice for days 0, 9–12, and 19). (e) The H2O2 levels in colons of mice at days 0 and 18–23 were measured using an Amplex Red assay. Apocynin (APO) was utilized as an NADPH oxidase inhibitor (–8 mice for day 0 and 4–7 for days 18–23). (f) Total bone marrow cells were isolated and MPO and Nox-2 expression was assessed using Western blot analysis. Representative images show bone marrow expression levels of MPO and Nox-2 in basal and colitic Eng +/+ and Eng +/− mice, with β-actin as the normalizing factor (days 0 and 18–23 images are derived from a single immunoblot). Histograms illustrate densitometric analysis of (g) MPO and (h) Nox-2 expression at all-time points tested (for MPO and Nox-2: mice for day 0 and 3–5 mice for days 7–9 and 18–23). versus corresponding day 0, versus corresponding Eng +/+ group, and versus corresponding untreated group. Results represent mean ± SEM.
