Journal: Scientific Reports

Article Title: Dietary polyamines promote intestinal adaptation in an experimental model of short bowel syndrome

doi: 10.1038/s41598-024-55258-4

Figure Lengend Snippet: Polyamines enhance mucosal defense factors in rats with massive intestinal resection. (A-B) Fecal and serum secretory IgA was measured using ELISA. n = 5–7/group. (C) IgA in ileum tissue was assessed using western blotting. n = 4–6/group. (D) Fecal mucin was measured using a fluorometric assay. n = 4–6/group. (E) Representative images of ileal villus showing immunostaining by Muc2 (original magnification, ×400; scale bars = 200 μm). Left graphs show the number of goblet cells per unit villous area and the size of goblet cell secretion granule. (F) The expression of Claudin-3 in the ileum tissue was measured by western blot analysis. Representative images of ileal villus showing immunostaining by Claudin-3 (original magnification, ×200; scale bars = 100 μm). (G) Serum DAO was measured by ELISA. n = 5–7/group. (H) Serum GLP-2 was measured by ELISA. n = 5–7/group. (I) Fecal short-chain fatty acid (SCFA) content was measured using high-performance liquid chromatography. n = 3–6/group. Data are presented as mean ± SD. Results of one-way ANOVA are represented as follows: * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Nonspecific antibody binding was blocked using 15% goat serum for 30 min. For staining of Ki-67, Muc2, and Claudin-3, the slides were then incubated with anti-Ki67 antibody (ab16667; Abcam, Cambridge, UK), anti-Muc2 rabbit monoclonal antibody (M01212-1; Boster Biological Technology, Pleasanton, CA, USA), and Claudin-3 polyclonal antibody (#PA5-32353; Invitrogen, Waltham, MA, USA) at 4°C overnight, respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Immunostaining, Expressing, High Performance Liquid Chromatography

Journal: Investigative Ophthalmology & Visual Science

Article Title: Ectoine Enhances Mucin Production Via Restoring IL-13/IFN-γ Balance in a Murine Dry Eye Model

doi: 10.1167/iovs.65.6.39

Figure Lengend Snippet: Antibodies Used in This Study

Article Snippet: MUC2 , CUSABIO , PA207869 , Rabbit , 1:3000.

Techniques:

Journal: Investigative Ophthalmology & Visual Science

Article Title: Ectoine Enhances Mucin Production Via Restoring IL-13/IFN-γ Balance in a Murine Dry Eye Model

doi: 10.1167/iovs.65.6.39

Figure Lengend Snippet: Mouse TaqMan Gene Expression Assays Used in This Study

Article Snippet: MUC2 , CUSABIO , PA207869 , Rabbit , 1:3000.

Techniques: Gene Expression

Journal: Investigative Ophthalmology & Visual Science

Article Title: Ectoine Enhances Mucin Production Via Restoring IL-13/IFN-γ Balance in a Murine Dry Eye Model

doi: 10.1167/iovs.65.6.39

Figure Lengend Snippet: Ectoine enhances production of secreted mucins in DS mice. ( A , C ) Representative images of IF staining showing the decreased production of MUC5AC ( A ) and MUC2 ( C ) in DS mice (DS+PBS) was recovered by ectoine eye drops (DS+Ect); Scale bars = 50 µm. The stitched images were made because the cornea and conjunctiva in some eyeball sections were too far apart to be captured on the same image. ( B , D ) IF intensity quantified the alternation of MUC5AC and MUC2 in 3 groups; each circle represents one eye. Data are shown as Mean ± SD, n = 6; the P values are shown in the graph as compared with the DS+PBS group.

Article Snippet: MUC2 , CUSABIO , PA207869 , Rabbit , 1:3000.

Techniques: Staining, Eye Drops

Journal: Investigative Ophthalmology & Visual Science

Article Title: Ectoine Enhances Mucin Production Via Restoring IL-13/IFN-γ Balance in a Murine Dry Eye Model

doi: 10.1167/iovs.65.6.39

Figure Lengend Snippet: Gene expression of MUC5AC, MUC2, MUC1, MUC4, MUC16 , and MUC15 by conjunctiva and cornea. ( A ) Gene expression of all 6 mucins in normal mice were detected at different mRNA levels based on the threshold cycle (Ct) value in real-time PCR with house-keeping gene GAPDH as an internal control. ( B ) The mRNA levels of all six mucins in conjunctiva of the three groups of mice. ( C ) The mRNA expression of MUC1, MUC4, and MUC16 by corneas in the three groups of mice. Data are shown as Mean ± SD, n = 6; the P values are shown in the graph as compared with the DS+PBS group.

Article Snippet: MUC2 , CUSABIO , PA207869 , Rabbit , 1:3000.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Control, Expressing

Journal: bioRxiv

Article Title: Phosphatase-independent suppression of mucosal inflammation and disease progression by Salmonella SopB

doi: 10.1101/2024.09.02.610793

Figure Lengend Snippet: (A) Small intestinal tissue sections stained with H&E of non-infected (n=3) neonates or neonates infected with wt (n=3) or Δ sopB (n=3) S . Typhimurium at day 2 p.i.. Bar=100 µm. (B) Quantification of the depth of the lamina propria as a measure of the tissue edema measured in 15-20 different areas of one tissue section per animal (non-infected neonates (n=3) or neonates infected with wt (n=3) or Δ sopB (n=3)). (C) Muc2 immunostaining (red) in small intestine tissue sections of non-infected (n=3) neonates and the neonates infected with wt (n=3) or Δ sopB (n=3) S . Typhimurium at day 2 p.i.. Counterstaining with WGA (white), DAPI (blue); autofluorescence (green). Bar=50 µm. (D) Percentage of goblet cells among intestinal epithelial cells in non-infected (n=3) neonates and neonates infected with wt (n=3) or Δ sopB (n=3) S . Typhimurium at day 2 p.i.. 10-20 images with the size of 312µm x 250µm were evaluated on one tissue section per animal. (E) Goblet cell size in tissue sections of uninfected (n=3) neonates and neonates infected with wt (n=3) or Δ sopB (n=3) S . Typhimurium at day 2 p.i.. 11-21 goblet cells (µm 2 ) were analyzed on each section. (F) TUNEL staining (red) of neonatal small intestine tissue sections of non-infected (n=2) neonates and neonates infected with wt (n=5) or Δ sopB (n=4) S . Typhimurium at day 2 p.i.. White boxes indicate enlarged images (i, ii). Counterstaining with DAPI (blue). Bar=50 µm; insert, 20 µm. (G) Number of TUNEL positive cells in tissue sections of uninfected (n=2) neonates and the neonates infected with wt (n=5) or Δ sopB (n=4) S . Typhimurium at day 2 p.i. 9 -12 images with the size of 624µm x 501µm were evaluated per animal. (H) Ki67 (red) immunostaining on small intestine tissue sections of the neonates infected with wt (n=3) or Δ sopB (n=4) S . Typhimurium at day 2 p.i.. Counterstaining with E-cadherin (green), WGA (white), and DAPI (blue). Bar=50 µm, insert: 20 µm. (I) Number of Ki67 positive cells in small intestinal tissue sections of uninfected (n=2) neonates and the neonates infected with wt (n=5) or Δ sopB (n=4) S . Typhimurium at day 2 p.i. 5-29 crypts were analyzed per section. Representative images are shown. Quantified data are shown as individual points in violin plots, solid lines represent the median. Data were analyzed using One-way ANOVA Kruskal–Wallis test with Dunn’s multiple comparisons post-test. ns, non-significant; *, p<0.05; ****, p<0.0001.

Article Snippet: Rabbit anti-Ki67 (Ab15580, Abcam), mouse anti-E-cadherin (610182, BD Transduction Laboratories), rat anti-PMN (Ly6-6B2, SeroTec), rabbit anti-Muc2 (GTX100664, BIOZOL) were used at the appropriate dilution followed by the fluorophore-conjugated donkey secondary antibody (Jackson ImmunoResearch).

Techniques: Staining, Infection, Immunostaining, TUNEL Assay