Fig 1: 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.
Fig 2: Enterocyte zonation is altered upon HFHSD.a, Activation of zonation scores along a pseudospatial ordering of EP and enterocytes from SI villi, which reflects the axis from villus bottom to tip. Cell scores for each zone were computed on reported markers18 and approximated by polynominal regression fits along the pseudospatial axis. Crossing points of the fitted lines define the partitioning into the five zones. b, Dot plots showing the expression of selected markers of enterocyte function in distal and proximal enterocytes. Cells are partitioned into zones along the inferred pseudospatial axis from villus bottom (zone 1) to tip (zone 5). c–e, Representative LSM images (c) and quantification of Fabp1 mean fluorescence intensity in ileal villi (d) and determination of the length of the Fabp1 positive zone (e). Scale bar, 50 µm, n = 3 CD mice and n = 4 HFHSD mice. Data are shown as mean ± s.d. of biologically independent samples. Statistical significance was determined by two-tailed Student’s t-test. f–h, Representative LSM images (f) and quantification of Apoa4 mean fluorescence intensity in ileal villi (g) and determination of the length of the Apoa4 positive zone (h). Scale bar, 50 µm, n = 3 CD mice and n = 4 HFHSD mice. Data are shown as mean ± s.d. of biologically independent samples. Statistical significance was determined by two-tailed Student’s t-test. i–k, Representative LSM images (i) and quantification of Apoa4 mean fluorescence intensity in duodenal villi (j) and determination of the length of the Apoa4 positive zone (k). Scale bar, 50 µm, n = 5 mice per group. Data are shown as mean ± s.e.m. of biologically independent samples. Statistical significance was determined by two-tailed Student’s t-test. Source data
Supplier Page from Abcam for Anti-liver FABP antibody [EPR20464]