Skip to main content
. 2019 Jun 10;8(3):407–426. doi: 10.1016/j.jcmgh.2019.06.001

Figure 5.

Figure 5

Validation of proximal small intestine markers that increase after SBR. (A) RNA FISH for Fabp1 shows significant upregulation of transcripts (fluorescein signal) per nucleated cell (DAPI) at days 7 and 70 after surgery. Day 7: n = 15 sham images, n = 15 SBR images (3 biological replicates). Day 70: n = 12 sham images, n =15 SBR images (3 biological replicates) (images acquired using Olympus FV1200 Confocal Microscope). (B) Immunohistochemistry staining for FABP1, FABP6, and SEPP1 in sham and SBR mice at postoperative day 7 shows qualitative increases in FABP1 and SEPP1, and decrease in FABP6, in SBR mice (images acquired using Nikon Eclipse 80i with Ds-Ri2 camera). (C) Immunofluorescence staining images for FABP1 (n = 12 sham images, n = 15 SBR images, 3 biological replicates), FABP6 (n = 18 sham images (4 biological replicates), n =11 SBR images (3 biological replicates)), and SEPP1 (n = 22 sham images (5 biological replicates), n = 18 SBR images (4 biological replicates)) were computationally analyzed to confirm significant protein-level changes corresponding to mRNA changes. White arrows indicate areas of intense SEPP1 expression in SBR (images acquired using Nikon Eclipse 80i with Ds-Ri2 camera). (D) Top to bottom: Western blot analysis of APOC3 in sham and SBR epithelial lysates, normalized to GAPDH and quantified (n = 3 sham and n = 4 SBR mice); qPCR validation of upregulated SBR genes Rbp2 and Ephx2 from SI tissue (n = 3 sham and n = 4 SBR mice). RNA-FISH images are at magnification 60×, scale bar = 30 μm. Immunohistochemistry stains are at 20×, scale bar = 100 μm. IF are at 40×, scale bar = 100 μm. All graphs are presented as mean ± SD. *P < .05, **P < .01, ****P < .0001.