Figure 1
From: Mouse and human Notch-1 regulate mucosal immune responses
Effect of Notch-1 knockdown (KD) on intestinal barrier function in vivo. (a) Claudin-5, -8, and occludin mRNA expression assessed in the distal colon of scrambled (Sc) or Notch-1 (N) small interfering RNA (siRNA)–treated wild-type (WT) mice by quantitative PCR (qPCR). (b) Claudin-5 immunofluorescence staining (red) in the distal colon of Sc or N siRNA-treated WT mice. The nuclei were stained with 4',6-diamidino-2-phenylindole (DAPI; blue). Samples were analyzed with a Leica SP5-DM Confocal microscope. These data are representative of three experiments. (c) Quantification of mean fluorescence intensity (MFI) per crypt associated with claudin-5 staining using the Adobe Photoshop software. (d) Assessment of changes in paracellular permeability for small molecules. The distal colon was harvested and the paracellular permeability was measured (assessed by flux measurements of dextran–fluorescein isothiocyanate (FITC) from the mucosal to serosal compartment, measured by spectrofluorometry). Data represent the mean±s.e.m. of at least 9 mice/group. P<0.05 was considered statistically significant.
