Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Dietary Stearic Acid Accelerates Intestinal Tumorigenesis via Fatty Acid-binding Protein 5 Without Promoting Obesity

doi: 10.1016/j.jcmgh.2026.101740

Figure Lengend Snippet: Spatial transcriptomics of the jejunum of mice fed the CD or SA-HFD. ( A ) UMAP of the small intestinal epithelium categorized into the enterocyte/crypt clusters. Each point represents a spot. ( B ) H&E staining of small intestinal sections analyzed using Visium ( upper panels ). The same sections are shown with spatial DimPlot visualization ( lower panels ). The colors of the spatial transcriptomics spots belonging uniquely to one factor were pink , green , and blue for enterocyte, crypt, and other, respectively. ( C ) Volcano plots comparing gene expression in Crypt clusters in mice fed the CD or SA-HFD. Red denotes upregulated genes with log2FC >0.6 and P value < .05, whereas blue denotes downregulated genes with log2FC <−0.6 and P value < .05. NS, not statistically significant. ( D ) KEGG pathway analysis of DEGs using ShinyGO. Enriched pathways in SA-HFD-fed mice compared with the findings in CD-fed mice are indicated (FDR <0.05). ( E ) Comparison of the cell-cycle and stem cell module in mice fed with CD or SA-HFD for 2 weeks. ( F ) Cell-cycle status indicated in stacked bar chart. ( G ) UMAP of the small intestinal epithelium categorized into 11 clusters ( upper panel ). Each point represents a spot. UMAP visualization highlights cluster 2 populations (stem cells, Paneth cells, and proliferating cells) in the small intestine of mice fed the CD or SA-HFD ( lower panel ). ( H ) The feature genes in cluster 2 were used as the marker gene for crypt cells, such as stem cells, Paneth cells, and proliferating cells. ( I ) The small intestine is presented using the spatial DimPlot. The colors of the spatial transcriptomics spots were uniquely assigned as pink for stem cells, Paneth cells, and proliferating cells. ( J ) KEGG pathway analysis of DEGs using ShinyGO. Enriched pathways in SA-HFD-fed mice compared with the findings in CD-fed mice are indicated (FDR <0.05). ( K ) The violin plots present Ppara, Ppard, Pparg, and Fgfbp1 expression in cluster 2 of CD- or SA-HFD-fed mice.

Article Snippet: The primary antibodies used in this study included Ki67 (GeneTex, #GTX16667), Lysozyme C (Santa Cruz, #sc-27958), FABP5 (R&D System, #AF1476), Chr-A (Santa Cruz, #sc-393941), Mucin-2 (SantaCruz, #sc-15334), DCAMKL1 (Abgent, #Ap7219b), CD44 (Biolegend, #103002), L-FABP (SantaCruz, #sc-374537), intestinal FABP (MyBioSource, #MBS178443), FABP6 (Proteintech, #13781-1-AP), Fgfbp1 (Proteintech, #25006-1-AP), and Msi-1 (eBioscience, #14-9896-82).

Techniques: Spatial Transcriptomics, Staining, Gene Expression, Comparison, Marker, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Dietary Stearic Acid Accelerates Intestinal Tumorigenesis via Fatty Acid-binding Protein 5 Without Promoting Obesity

doi: 10.1016/j.jcmgh.2026.101740

Figure Lengend Snippet: SA induced FABP5 expression in intestinal mucosa and organoids. ( A ) Representative images of jejunal sections stained with anti-FABP1, anti-FABP2, anti-FABP5, or anti-FABP6. The jejunum was obtained from C57BL/6 mice fed chow diet. The right panels present a higher-magnification view of the crypt area in the left panels ( box ). The arrowheads indicate FABP5-positive cells. The scale bars represent 20 μm ( right ) or 100 μm ( left ). ( B ) Representative fluorescent images of jejunal sections co-stained with anti-FABP5 ( green ), anti-Chr-A ( red ), anti-Ki67 ( orange ), and DAPI ( blue ). The jejunum was obtained from C57BL/6 mice fed chow diet. The right upper and lower panels present a higher-magnification view of the area surrounded by white and yellow dashed lines in the left panel , respectively. FABP5 merged with Chr-A or Ki67 signals appear yellow . The scale bars represent 20 μm. ( C ) A representative fluorescent image of jejunal sections co-stained with anti-FABP5 ( green ), anti-GFP ( red ), and DAPI (blue). The jejunum was obtained from Lgr5-EGFP reporter mice fed chow diet. The scale bars represent 10 μm. ( D ) A representative fluorescent image of jejunal sections from Bmi1-GFP reporter mice fed chow diet ( left ). Bmi1 ( green ) was visualized by nuclear staining ( blue ). Serial sections were stained with anti-FABP5 and DAPI ( blue; right ). Bmi1-GFP+ cells ( white arrowhead ) and FABP5+ cells ( white arrow ) are denoted. The scale bars represent 10 μm. ( E ) Representative fluorescent image of jejunal sections co-stained with anti-FABP5 ( green ), anti-Fgfbp1 ( red ), and DAPI ( blue ). The jejunum was obtained from C57BL/6 mice fed the chow diet. Yellow arrowheads indicate FABP5+ Fgfbp1+ cells. The scale bars represent 20 μm. ( F ) Representative fluorescent image of jejunal sections co-stained with anti-CD44 ( green ), anti-Fgfbp1 ( red ), and DAPI ( blue ). The jejunum was obtained from C57BL/6 mice fed the chow diet. The scale bars represent 20 μm. ( G ) A representative fluorescent image of jejunal sections co-stained with anti-FABP5 ( green ), anti-Lysozyme C ( red ), and DAPI ( blue ). The jejunum was obtained from C57BL/6 mice fed the chow diet. The scale bars represent 10 μm. ( H ) Representative images of organoids derived from FABP5+ cells isolated from the jejunum and ileum of FABP5 reporter mice. Organoids were cultured in basal medium containing growth factors. Representative images of spheroids ( left ) and organoids ( right ) are presented. The data are representative of 2 separate experiments (n = 2), which gave similar results. The scale bars represent 50 μm ( right ) or 200 μm ( left ). ( I ) Representative fluorescent image of organoids derived from FABP5+ cells stained with anti-Lysozyme C ( green ), anti-Ki67 ( magenta ), and DAPI ( blue ). The scale bars represent 10 μm. ( J ) Representative fluorescent images of jejunal sections co-stained with anti-FABP5 ( red ) and anti-CD34 ( green; upper panels ) or anti-CD31 ( green; lower ). The jejunum was obtained from C57BL/6 mice fed chow diet. DAPI ( blue; right panels ) was used for nuclear staining. FABP5 signals merged with CD34 or CD31 signals appear yellow . The scale bars represent 20 μm. ( K ) Relative mRNA expression of Fabp1, Fabp2, and Fabp6 in epithelial cells isolated from the jejunum of C57BL/6 mice fed the CD, LA-HFD, or SA-HFD for 2 weeks (each group, n = 8). ( L ) Violin plots of Fabp5 in cluster 2. ( M ) Representative immunohistochemical images of the jejunum stained with anti-FABP5. The jejunum was obtained from C57BL/6 mice fed the CD ( left panels ), LA-HFD ( middle panels ), or SA-HFD ( right panels ) for 2 weeks. The numbers of FABP5+ cells per crypt were counted. The data represent the average counts in 8 to 15 crypts from individual mice (n = 4). The scale bars represent 20 μm. ( N ) Representative fluorescent images of the jejunum co-stained with anti-FABP5 ( red ), anti-Ki67 ( green ), and DAPI ( blue ). The jejunum was obtained from C57BL/6 mice fed the CD ( left panels ), LA-HFD ( middle panels ), or SA-HFD ( right panels ) for 2 weeks. The numbers of Ki67+ FABP5+ cells and Ki67- FABP5+ cells per crypt were counted. The data represent the average counts in 8 to 15 crypts from individual mice (n = 4). The scale bars represent 20 μm. ( O ) Representative fluorescent images of untreated or fatty acid-treated murine organoids stained with anti-FABP5 ( green ) and DAPI ( blue ). Quantification of the area of FABP5+ cells in organoids normalized to the DAPI-positive area. The scale bars represent 50 μm. ( P ) Representative images of formalin-fixed, paraffin-embedded sections of human small intestine stained with anti-FABP5. Representative images of surgical specimens of the small intestine obtained from 4 patients. The scale bars represent 50 μm. ( Q ) Relative mRNA expression of FABP5 and Fgfbp1 in normal colon (n = 31) and colon cancer (n = 31) tissues from the same patients. Data were retrieved from the NCBI GEO database ( GSE8671 ). ( R ) Cell growth assay. The data represent the mean ± SEM. ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001.

Article Snippet: The primary antibodies used in this study included Ki67 (GeneTex, #GTX16667), Lysozyme C (Santa Cruz, #sc-27958), FABP5 (R&D System, #AF1476), Chr-A (Santa Cruz, #sc-393941), Mucin-2 (SantaCruz, #sc-15334), DCAMKL1 (Abgent, #Ap7219b), CD44 (Biolegend, #103002), L-FABP (SantaCruz, #sc-374537), intestinal FABP (MyBioSource, #MBS178443), FABP6 (Proteintech, #13781-1-AP), Fgfbp1 (Proteintech, #25006-1-AP), and Msi-1 (eBioscience, #14-9896-82).

Techniques: Expressing, Staining, Derivative Assay, Isolation, Cell Culture, Immunohistochemical staining, Formalin-fixed Paraffin-Embedded, Growth Assay

Journal: Nutrients

Article Title: Neuroprotective Effects of Desert Milk Exosomes in LPS-Induced Cognitive Decline: Role of Microglial M2 Polarization and AMPK Signaling

doi: 10.3390/nu18020315

Figure Lengend Snippet: Quality assessment of proteome data by label-free quantification. ( A ) Principal component analysis. ( B ) Hierarchical clustering analysis of differential protein expression profiles between D-Exo and ND-Exo. Red = Exosomes proteins with higher expression, green = Exosomes proteins with lower expression. ( C ) Difference expression pattern between D-Exo and ND-Exo. FDR = false discovery rate; FC = fold change. ( D ) The SIL1, FN1 protein with representative image by WB. ( E , F ) Expression of SIL1 and FN1 proteins in D-Exo and ND-Exo by WB method. Data are presented as mean ± SEM ( n = 3 per group). Statistically significant differences were indicated: * p < 0.05, ** p < 0.01, *** p < 0.001, ns > 0.05.

Article Snippet: Following blocking with 5% BSA, the membranes were incubated overnight at 4 °C with the primary antibody for FN1 (cat. #15613-1-AP, Proteintech, Shanghai, China), SIL1 (cat. #24110-1-AP, Proteintech, Shanghai, China), AMPK (cat. #10929-2-AP, Proteintech, Shanghai, China), phospho-AMPK (cat. #80209-6-RR, Proteintech, Shanghai, China), and GAPDH (cat. #10494-1-AP, Proteintech, Shanghai, China).

Techniques: Quantitative Proteomics, Expressing

Journal: PLoS ONE

Article Title: Analysis of the Genome of a Korean Isolate of the Pieris rapae Granulovirus Enabled by Its Separation from Total Host Genomic DNA by Pulse-Field Electrophoresis

doi: 10.1371/journal.pone.0084183

Figure Lengend Snippet: Genomic DNA or BAC DNA isolation and purification was followed by size fractionation and ligation into a pUC118 ready vector for 4 o C followed with transformation by electroporation into DH5α. The quality of thus constructed shotgun library was checked by titering (40 µl of cell stock, white: blue = 400∶100). The number of clones was approximately 20,000 in total. 96 clones were selected and sequenced including insert size check, E. coli and vector % check.

Article Snippet: Equivalent volumes of fosmid DNA clones were digested with Not I to obtain 3-7 kb DNA pieces that were then ligated into a purified pUC118 Bam HI/BAP ready vector (Takara Bio Inc., Shiga, Japan) .

Techniques: DNA Extraction, Purification, Fractionation, Ligation, Plasmid Preparation, Transformation Assay, Electroporation, Construct, Clone Assay