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. 2013 May 6;110(23):9451–9456. doi: 10.1073/pnas.1219997110

Fig. 4.

Fig. 4.

The severity of experimental NEC is increased in eNOS−/− mice and reduced after administration of sildenafil, and TLR4 reduces the expression of eNOS in isolated MIMECs in a MyD88-dependent manner. (A and B) Photographs of the intestines and histomicrographs obtained from eNOS−/− (A) or wild-type mice that had been administered sildenafil as described in Materials and Methods (B) that were either breast fed (i and ii) or induced to develop NEC (iii and iv). (C) qRT-PCR showing expression of IL-6 within the intestinal mucosa (i) or NEC severity score (ii) in wild-type or eNOS−/− mice under the condition indicated. *P < 0.05 wild-type NEC vs. breast fed; **P < 0.05 NEC plus sildenafil vs. NEC; ***NEC in eNOS−/− vs. NEC in wild-type mice. Results are representative of three separate experiments with at least 10 mice per experiment. (D and E) Mouse MIMECs were isolated from the ileum of mice based upon their staining for CD31+ CD45−. Mice had been treated 6 h prior with saline or LPS and were wild type, TLR4−/−, TLR4Δendoth, MyD88−/−, or TRIF−/− or had been pretreated with the NF-κB inhibitor BAY 11 as indicated. (D) Representative plot of sorted MIMECs in the presence of saline or LPS as shown. (E) Percent eNOS-positive MIMECs from the indicated strain treated with saline or LPS as indicated. *P < 0.05. Results are representative of three separate experiments. All values are mean ± SEM.