vegfc  (Santa Cruz Biotechnology)

Journal: Journal of Advanced Research

Article Title: Fish oil reduces intestinal fat absorption by promoting lacteal junction zippering via GPR120-VEGFR3-MLCK pathway

doi: 10.1016/j.jare.2025.05.026

Figure Lengend Snippet: Fish oil gavage suppressed intestinal fat absorption associated with elevated proportion of lacteal zipper-like junctions and decreased expression of VEGFR3 in mice jejunum. (A) Oral fat tolerance test plasma TG level schematic. (B) Fat tolerance test serum TG levels at indicated times. (C, D) Effects of lard oil (with/without fish oil) on the expression of VEGFC, VEGFR3, and chylomicron formation related protein (ApoB48, MTTP) in the jejunum of chow diet-fed mice. (E) Button-like junctions (open) allow chylomicrons into lacteals; zipper-like junctions (closed) prevent this. (F, G) Morphometric analysis of jejunal villus length and lacteal dimensions (length/width) in mice: Representative imaging and quantitative comparisons. (H, I) Visual and comparative analysis of VE-cadherin positive junctions in mice jejunum lacteals. Each boxed area is highly magnified to differentiate button-like (red arrowheads) and zipper-like (yellow arrowheads) junctions. Scale bars, 50 μm. Data are represented as means ± SEM. Two-tailed unpaired Student’s t -test was performed to compare lard oil (with/without fish oil)-treated mice versus at the same time points. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Primary antibodies include CD36 (proteintech,18836–1-AP), VEGFC (proteintech,67116–1-Ig), VEGFR3 (R&D, AF743), ApoB48 (ThermoFisher, MA5-35458), MTTP (ThermoFisher, PA5-102474,), MLCK (ThermoFisher Scientific, PA5100908), GPR120 (Santa Cruz, sc-9901), p-MLC2 (CST, 95777S, Thr18/Ser19), GAPDH (Bioworld, AP0063), β-actin (Bioworld, AP0060) and β-Tubulin (Bioworld, AP0064).

Techniques: Expressing, Clinical Proteomics, Imaging, Two Tailed Test

Journal: Journal of Advanced Research

Article Title: Fish oil reduces intestinal fat absorption by promoting lacteal junction zippering via GPR120-VEGFR3-MLCK pathway

doi: 10.1016/j.jare.2025.05.026

Figure Lengend Snippet: DHA and EPA increased the proportion of zipper-like junctions via downregulation of VEGFR3 signaling in palmitic acid (PA)-treated LECs. (A-D) Effects of DHA and EPA (20 μM) on the proportion of zipper-like junctions (A, B) and the expression of VEGFR3 (C, D) in PA (100 μM)-treated LECs. (E, F) Effects of VEGFC (100 ng/mL)-induced VEGFR3 activation on the proportion of zipper-like junctions in PA (100 μM)-treated LECs. White dashed line: VE-cadherin negative junctions; white arrowhead: button-like junctions; yellow arrowhead: zipper-like junctions. Data are represented as means ± SEM. Two-tailed unpaired Student’s t -test was performed to compare PA versus PA + DHA/EPA, PA + VEGFC versus PA + DHA/EPA + VEGFC-treated groups at the same time points. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Primary antibodies include CD36 (proteintech,18836–1-AP), VEGFC (proteintech,67116–1-Ig), VEGFR3 (R&D, AF743), ApoB48 (ThermoFisher, MA5-35458), MTTP (ThermoFisher, PA5-102474,), MLCK (ThermoFisher Scientific, PA5100908), GPR120 (Santa Cruz, sc-9901), p-MLC2 (CST, 95777S, Thr18/Ser19), GAPDH (Bioworld, AP0063), β-actin (Bioworld, AP0060) and β-Tubulin (Bioworld, AP0064).

Techniques: Expressing, Activation Assay, Two Tailed Test

Journal: Journal of Advanced Research

Article Title: Fish oil reduces intestinal fat absorption by promoting lacteal junction zippering via GPR120-VEGFR3-MLCK pathway

doi: 10.1016/j.jare.2025.05.026

Figure Lengend Snippet: DHA and EPA inhibited MLCK signaling in a GPR120-VEGFR3 dependent manner in PA-treated LECs. (A, B) Effects of DHA and EPA (20 μM) on the expression of MLCK and p-MLC 2 in PA (100 μM)-treated LECs. (C, D) Effects of AH-7614 (50 μM) treatment on the expression of GPR120, MLCK and p-MLC 2 in PA (100 μM)-treated LECs. (E, F) Effects of VEGFC (100 ng/mL)-induced VEGFR3 activation on the expression of VEGFR3, MLCK and p-MLC 2 in PA (100 μM)-treated LECs. Two-tailed unpaired Student’s t -test was performed to compare PA versus PA + DHA/EPA-treated groups at the same time points.

Article Snippet: Primary antibodies include CD36 (proteintech,18836–1-AP), VEGFC (proteintech,67116–1-Ig), VEGFR3 (R&D, AF743), ApoB48 (ThermoFisher, MA5-35458), MTTP (ThermoFisher, PA5-102474,), MLCK (ThermoFisher Scientific, PA5100908), GPR120 (Santa Cruz, sc-9901), p-MLC2 (CST, 95777S, Thr18/Ser19), GAPDH (Bioworld, AP0063), β-actin (Bioworld, AP0060) and β-Tubulin (Bioworld, AP0064).

Techniques: Expressing, Activation Assay, Two Tailed Test

Journal: Investigative Ophthalmology & Visual Science

Article Title: Single-Cell Transcriptomics Shows Cellular Heterogeneity, Intercellular Communication, and Extracellular Matrix Remodeling in Corneal Fibrosis In Vivo

doi: 10.1167/iovs.66.13.48

Figure Lengend Snippet: Intercellular communication within ECM-remodeling subclusters. (a) Network of inferred ligand–receptor exchanges among four stromal subclusters: 0-quiescent keratocytes, 1-activated keratocytes/fibroblasts, 2-progenitor-like keratocytes, and 3-proliferative fibroblasts/myofibroblasts; node size indicates the number of cells, and edge width denotes total signaling strength. (b) Heatmap of pathway-level enrichment across the four ECM-remodeling subclusters. (c–j) Chord diagrams with accompanying dot plots showing a representative subset of pathways; MIF (c) , NECTIN (d) , HGF (e) , POSTN (f) , NAMPT (g) , TWEAK/TNFSF12 (h) , EPHA (i) , and VEGF (j) , selected from the heatmap on the basis of differential expression and to cover distinct signaling families. The representative ligand–receptor pairs include MIF–CD74+CD44 or ACKR3, NECTIN3–NECTIN1/2, HGF–MET, POSTN–ITGAV+ITGB5, NAMPT–INSR or ITGA5+ITGB1, TNFSF12–TNFRSF12A, EFNA1/5–EPHA2/4/5, and VEGFA/VEGFC–VEGFR2/FLT4. The dot size reflects interaction strength, and the color intensity indicates communication probability.

Article Snippet: For immunofluorescence, the tissue sections were deparaffinized, subjected to citrate-based antigen retrieval (pH 6.0, 95°C, 20 minutes), blocked in 5% BSA, and incubated overnight at 4°C with primary antibodies targeting the following proteins: α-smooth muscle actin (α-SMA) (1:300; Abcam Inc., Waltham, MA, USA; Cat no. AB322917 ), PDGFA (1:100; Santa Cruz Biotechnology, Santa Cruz, CA, USA; Cat no. sc-9974), vascular endothelial growth factor-A (VEGFA) (1:100; Santa Cruz Biotechnology; Cat no. sc-365578), transforming growth factor-β1 (TGFβ1) (1:100; Santa Cruz Biotechnology; Cat no. sc-130348), TGFβ2 (1:100; Santa Cruz Biotechnology; Cat no. sc-374659), ephrin type-A receptor-5 (1:100; Santa Cruz Biotechnology; Cat no. sc-517242), VEGFC (1:100; Santa Cruz Biotechnology; Cat no. sc-374628), nectin cell adhesion molecule-3 (1:100; Santa Cruz Biotechnology; Cat no. sc-271611), collagen type-I α1 chain (COL1A1) (1:100; Santa Cruz Biotechnology; Cat no. sc-293182), junctional adhesion molecule-A (JAMA) (1:100; Santa Cruz Biotechnology; Cat no. sc-53623), macrophage migration inhibitory factor (MIF) (1:100; Santa Cruz Biotechnology; Cat no. sc-271631), LAMININ5 (1:100; Santa Cruz Biotechnology; Cat no. sc-13587), hepatocyte growth factor-α chain (HGFα) (1:100; Santa Cruz Biotechnology; Cat no. sc-374422), TNF-like weak inducer of apoptosis (TWEAK) (1:100; Santa Cruz Biotechnology; Cat no. sc-56248), nicotinamide phosphoribosyltransferase (NAMPT) (1:100; Santa Cruz Biotechnology; Cat no. sc-393444), and periostin (POSTN) (1:100; Santa Cruz Biotechnology; Cat no. sc-398631).

Techniques: Quantitative Proteomics