Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Inhibition of BMP2 and BMP4 Represses Barrett’s Esophagus While Enhancing the Regeneration of Squamous Epithelium in Preclinical Models

doi: 10.1016/j.jcmgh.2023.01.003

Figure Lengend Snippet: (A) Clustering of RNA expression in biopsies from Barrett’s and squamous tissue and from a validation cohort (Wang 2013, GSE26886).27 Gene-clustered heatmap was drawn for quantitative PCR data using the heatmap package, after log transformation and calculation of z-scores for all samples per gene. (B) Quantitative PCR of SHH/WNT pathway genes in biopsies from Barrett’s and squamous tissue from AMC patients (left) and from a validation cohort (right) (Wang 2013, GSE26886).27 Genes significantly differentially expressed in Barrett’s compared with squamous in the AMC cohort (P < .05). (C) Western blot analysis of BMP2, BMP4, BMP5, and BMP6 in Barrett patient’s (BE) biopsies (left panel). Levels of BMPs in Barrett’s biopsies were quantified and normalized to squamous tissue (SQ). Data are represented in box and whiskers plot of at least 4 independent experiments. (D) BMP2, BMP4, BMP7, and SHH mRNA levels by quantitative PCR in mouse esophageal keratinocytes, human normal squamous esophageal EPC2-hTERT, and Barrett’s CP-A cells, after treatment with glycine-conjugated bile acid (100 μmol/L) for 8, 24, or 48 hours. Data are represented in box and whiskers plots as mean of at least 3 independent experiments. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001. ns: not significant.

Article Snippet: The membranes were blocked with blocking solution (5% nonfat milk in Tris-buffered saline with 1% Tween-20) and afterwards incubated with the appropriate primary antibody solution anti-BMP2 (Peprotech), anti-BMP4 (R&D), anti-BMP5 (Abcam), and anti-BMP6 (R&D) at a 1:500 dilution in blocking solution overnight at 4°C.

Techniques: RNA Expression, Real-time Polymerase Chain Reaction, Transformation Assay, Western Blot

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Inhibition of BMP2 and BMP4 Represses Barrett’s Esophagus While Enhancing the Regeneration of Squamous Epithelium in Preclinical Models

doi: 10.1016/j.jcmgh.2023.01.003

Figure Lengend Snippet: List of the Primers (Sense and Antisense) Used for Quantitative PCR

Article Snippet: The membranes were blocked with blocking solution (5% nonfat milk in Tris-buffered saline with 1% Tween-20) and afterwards incubated with the appropriate primary antibody solution anti-BMP2 (Peprotech), anti-BMP4 (R&D), anti-BMP5 (Abcam), and anti-BMP6 (R&D) at a 1:500 dilution in blocking solution overnight at 4°C.

Techniques:

Journal: Haematologica

Article Title: Iron overload induces BMP6 expression in the liver but not in the duodenum

doi: 10.3324/haematol.2010.031963

Figure Lengend Snippet: Effect of dietary iron-enrichment or Hfe-deficiency on hepatic iron concentrations and Bmp6 gene expression in the liver and the duodenum of 7-week old C57BL/6, DBA/2, and 129/Sv mice. (A) Increase in hepatic iron concentrations and expression ratios of Bmp6 transcripts normalized to the Hprt reference gene mRNA (and 95% confidence intervals) in wild-type animals fed an iron-enriched diet versus animals fed an iron-balanced diet (5 mice per group). (B) Increase in hepatic iron concentrations and expression ratios of Bmp6 transcripts normalized to the Hprt reference gene mRNA (and 95% confidence intervals) in Hfe-deficient versus wild-type mice (5–6 mice per group).

Article Snippet: Similar results were obtained with the Santa Cruz N-19 anti-BMP6 antibody ( data not shown ).

Techniques: Gene Expression, Expressing

Journal: Haematologica

Article Title: Iron overload induces BMP6 expression in the liver but not in the duodenum

doi: 10.3324/haematol.2010.031963

Figure Lengend Snippet: Cellular localization of iron and BMP6 in the liver and the duodenum of mice with iron overload secondary to an iron-enriched diet or due to Hfe-deficiency. Iron deposits are vizualized by Perls’ staining (1). BMP6 expression was detected by immunohistochemistry in the liver (2) and the duodenum (3). As expected, although Bmp6-deficient mice (A) have the highest iron accumulation, no Bmp6 was detected. C57BL/6 mice with secondary iron overload (B) and DBA/2 Hfe-deficient mice (C) have significant amounts of liver iron and Bmp6 in their liver but not in their duodenum. 129/Sv mice with secondary iron overload (D) have the lowest amount of liver iron and this correlates with Bmp6 staining in the liver. Original magnification x100 (1) or x200 (2 and 3).

Article Snippet: Similar results were obtained with the Santa Cruz N-19 anti-BMP6 antibody ( data not shown ).

Techniques: Staining, Expressing, Immunohistochemistry