Loss of Nrp1 in SMC affects the expression of markers of contractile phenotype.

(A) qPCR analysis of SMC contractile phenotype markers, smMHC (n = 5) and Sm22-alpha (n≥3), on colonic samples from mice ≥16 months old: means±SD; *P<.05, **P<.005, ***P = .0005. (B) smMHC/SMA immunohistochemistry in colonic tissue from ≥16 months old mice. (C) Semi-quantitative RT-PCR for the SMA and SMB smMHC isoforms in whole colon cDNA from ≥16 month-old mice. Representative results are shown from 3 mice (Nrp1^fl/+, n = 4; Nrp1^+/−, n = 6; Nrp1^SMKO, n = 6; *P = .04). (D) Immunofluorescent staining of SK3 and SMA in colons of ≥16 months old mice shows enhanced expression of SK3 in the colonic SMC (arrows) of Nrp1^SMKO compared to Nrp1^+/− controls. (E) Western blot of SK3 expression in colon lysates from ≥16 month-old mice. Densitometric quantification of SK3 was normalised to beta-actin expression: means±SD; *P = .0233, n = 4.