Journal: PLoS ONE

Article Title: Efficient Assessment of Developmental, Surgical and Pathological Lymphangiogenesis Using a Lymphatic Reporter Mouse and Its Embryonic Stem Cells

doi: 10.1371/journal.pone.0157126

Figure Lengend Snippet: (A-J) Embryonic tissues: bright field (A) and fluorescence (B) images of the Prox1-tdTomato embryo (E17.5). Distinct lymphatic networks shown in the embryonic skin (C-E). (E) Enlarged image of the boxed area in panel D. Lymphatic vessels in the embryonic liver (F,G) and mesentery membrane (H). Note that hepatocytes (I) and tail nerves (J) are also positive for tdTomato. (K-Y) Adult tissues: headshots of adult wild type and Prox1-tdTomato transgenic mice were taken under a bright light (K), bright and fluorescent light (L) and fluorescent light (M). Lymphatic vessels were easily detectable in various tissues such as the ear (N), eye (O,P), tail (Q), tongue (R), trachea (S), diaphragm muscle (T), bladder (U,V), intestine (W), mesentery (X) and lymph node (Y). Corneal limbal lymphatic (arrow) and Schlemm’s canal (arrowhead) of the eye were shown in two consecutive focal planes (O,P). Bilateral lymphatic collectors in the tail were marked with two arrows (Q).

Article Snippet: Prox1 staining was performed using anti-Prox1 antibody (R&D Systems).

Techniques: Fluorescence, Membrane, Transgenic Assay

Journal: PLoS ONE

Article Title: Efficient Assessment of Developmental, Surgical and Pathological Lymphangiogenesis Using a Lymphatic Reporter Mouse and Its Embryonic Stem Cells

doi: 10.1371/journal.pone.0157126

Figure Lengend Snippet: (A-H) Axillary lymphadenectomy model: Fluorescent light enables the researchers to easily locate the axillary lymph node (A-D). From a different set of mice, the left-side axillary lymph nodes were removed, while the right-side nodes were left intact. To enhance lymphatic growth in the surgical site, 9-cis RA was treated for the first 5 days after surgery. After 4 weeks, lymphatic vessels were visualized in the left vs . right side of the armpit of the mice with asterisks indicating the original locations of the lymph nodes (E,F). Images in the boxed areas (E, F) are shown in panels G and H. (I-Q) Lymph node transplantation model: the axillary lymph node in Prox1-tdTomato mice (recipient) was replaced with the matching lymph node from Prox1-EGFP mice (donor), followed by administration of 9-cis RA for 4 weeks. Panels O, P and Q are enlarged images of the boxed areas in panels K, M and N, respectively. (R-U) Tail lymphedema model: representative images of the circumcised tails at post-operational days 1 and 15 are shown (R,T). At both days, the circumcised tail skins were prepared inside-out to visualize dermal lymphatic vessels (S,U). Asterisks mark the wounded area and black dotted lines indicate the wound border. Note the numerous newly formed lymphatics invading into the healed area (S).

Article Snippet: Prox1 staining was performed using anti-Prox1 antibody (R&D Systems).

Techniques: Transplantation Assay

Journal: PLoS ONE

Article Title: Efficient Assessment of Developmental, Surgical and Pathological Lymphangiogenesis Using a Lymphatic Reporter Mouse and Its Embryonic Stem Cells

doi: 10.1371/journal.pone.0157126

Figure Lengend Snippet: (A-E) Bladder lymphangiogenesis model: Adult Prox1-tdTomato mice were i.p. injected with phosphate-buffered saline (A,C) or cyclophosphamide (B,D) at days 1 and 4. At day 9, lymphatic vessels in their bladders were visualized. Morphometric analyses revealed a significantly increase in the number of branch (marked with arrowhead), loop (arrow) and blind ends (double arrowhead) of lymphatic vessels in the CYP-treated bladder compared to the PBS-treated control (CTR) bladder (E). (F-J) Tumor implantation model: GFP-labeled tumor cells were implanted into the skin of immunodeficient athymic Prox1-tdTomato mouse, generated by crossing athymic nude mice and Prox1-tdTomato mice. Images of the GFP-labeled tumor mass (F), dermal lymphatic in the surrounding tissues (G) and their merged image (H) are shown. (I) and (J) are enlarged images of the boxed areas in panel (H) and (I), respectively. Arrows mark luminal valves.

Article Snippet: Prox1 staining was performed using anti-Prox1 antibody (R&D Systems).

Techniques: Injection, Saline, Control, Tumor Implantation, Labeling, Generated

Journal: PLoS ONE

Article Title: Efficient Assessment of Developmental, Surgical and Pathological Lymphangiogenesis Using a Lymphatic Reporter Mouse and Its Embryonic Stem Cells

doi: 10.1371/journal.pone.0157126

Figure Lengend Snippet: (A,B) Bright field (BF) and fluorescent (UV) images of the head of Prox1-tdTomato, Prox1-tdTomato/Prox1-EGFP and Prox1-EGFP lymphatic-specific reporter mice. (C-K) Expressions of tdTomato and/or EGFP proteins were visualized in the skin of embryos (EM, C-E) and new pups (P1, F-H), or in the bladder of adult (BL, I-J) of each genotype. (L-N) Prox1 whole-mount staining of the ear of tdTomato adult mouse.

Article Snippet: Prox1 staining was performed using anti-Prox1 antibody (R&D Systems).

Techniques: Staining

Journal: Cell reports

Article Title: Early Seizure Activity Accelerates Depletion of Hippocampal Neural Stem Cells and Impairs Spatial Discrimination in an Alzheimer’s Disease Model

doi: 10.1016/j.celrep.2019.05.101

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse anti-Prox1 , PhosphoSolutions , Cat#: 1685-Prox1; RRID: AB_2492217.

Techniques: Recombinant, Imaging, Software, Microscopy

Journal: Cells

Article Title: Conjugated Bile Acids Promote Lymphangiogenesis by Modulation of the Reactive Oxygen Species–p90RSK–Vascular Endothelial Growth Factor Receptor 3 Pathway

doi: 10.3390/cells12040526

Figure Lengend Snippet: Antibodies used in Western blot, immunohistochemistry, and immunofluorescence.

Article Snippet: Prox1 , IP, WB , 1:1000 , Cell Signaling Technology , 14963 , AB_2783562.

Techniques: Western Blot, Immunohistochemistry, Immunofluorescence

Journal: Cells

Article Title: Conjugated Bile Acids Promote Lymphangiogenesis by Modulation of the Reactive Oxygen Species–p90RSK–Vascular Endothelial Growth Factor Receptor 3 Pathway

doi: 10.3390/cells12040526

Figure Lengend Snippet: Conjugated BAs promote ROS production and induces p90RSK-mediated Prox1 SUMOylation and enhances VEGFR3. TCA (100 µM) induced ( A ) cellular ROS production at 45 min and ( B ) mtROS production after 12 h of treatment in HLEC, measured by fluorometric assay in plate reader. TGR5 antagonist SBI-115 (10 µM) pre-treatment for 1 h inhibited the TCA-induced mtROS production. ( C ) Western blot analysis showed that TCA (100 µM)-induced p90RSK phosphorylation in HLECs after 24 h, which was inhibited by p90RSK inhibitor BI-D1870. ( D ) After 24 h treatment with TCA (100 µM), the Prox1 was immunoprecipitated using anti-Prox1 antibody and probed with anti-SUMO1 antibody. TCA treatment induced Prox1 SUMOylation in HLECs which was inhibited by inhibition of p90RSK activation by using BI-D1870. ( E ) TCA (100 µM) induced VEGFR3 mRNA expression which was inhibited by p90RSK inhibitor BI-D1870 and ROS inhibitor NAC. ( F ) Prox1 gene was knocked down in HLECs by siProx1 and ( G ) Prox1 knockdown inhibited the TCA-induced VEGFR3 expression. ( H ) TCA induced tube formation of HLECs in Matrigel at 4 h, which was inhibited by the TGR5 antagonist SBI-115, p90RSK inhibitor BI-D1870. ( I ) TCA (100 µM) induced the HLEC invasion in 3D collagen matrix at 20 h and was inhibited by BI-D1870 or TGR5 antagonist SBI-115. Values represent mean ± SD, n ≥ 3. Statistical analysis was done by ANOVA followed by Fishers LSD. *, **, ***, **** indicates p ≤ 0.05, p ≤ 0.01, p ≤ 0.001, p ≤ 0.0001, respectively, as compared to respective control., $$$, $$$$ indicates p ≤ 0.001, p ≤ 0.0001, respectively, as compared to TCA. #, ###, #### indicates p ≤ 0.05, p ≤ 0.001, p ≤ 0.0001, respectively, as compared to TCDCA.

Article Snippet: Prox1 , IP, WB , 1:1000 , Cell Signaling Technology , 14963 , AB_2783562.

Techniques: Western Blot, Phospho-proteomics, Immunoprecipitation, Inhibition, Activation Assay, Expressing, Knockdown, Control

Journal: Cells

Article Title: Conjugated Bile Acids Promote Lymphangiogenesis by Modulation of the Reactive Oxygen Species–p90RSK–Vascular Endothelial Growth Factor Receptor 3 Pathway

doi: 10.3390/cells12040526

Figure Lengend Snippet: Schematic showing increased conjugated BAs induce pathological lymphangiogenesis through an ROS-p90RSK-YAP-VEGFR3-dependent mechanism. Conjugated BAs bind to TGR5 expressed by the LECs and activate several key members of the redox or oxidative stress pathway. The activation of the redox pathway in turn produces ROS and activates the p90RSK via its phosphorylation at S380. Activated pp90RSK induced the SUMOylation of Prox1, and SUMOylated Prox1 increased the transcription of VEGFR3, ultimately increasing lymphangiogenesis. Conjugated BA-induced ROS also activated the YAP, which in turn increased the lymphangiogenesis. BA also enhanced the cellular metabolism by increasing the mRNA expression of the metabolic genes, namely PFKP , HK2 , CO I , and FASN , that play roles in enhancing lymphangiogenesis. Generated with BioRender.com (Toronto, ON, Canada).

Article Snippet: Prox1 , IP, WB , 1:1000 , Cell Signaling Technology , 14963 , AB_2783562.

Techniques: Activation Assay, Phospho-proteomics, Expressing, Generated