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Fish oil gavage suppressed intestinal fat absorption associated with elevated proportion of lacteal zipper-like junctions and decreased expression <t>of</t> <t>VEGFR3</t> 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 <t>VEGFC,</t> 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.)
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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.)

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

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.)

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

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.

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