Journal: International Journal of Inflammation

Article Title: Interleukin-1 β Expression Is Required for Lysophosphatidic Acid-Induced Lymphangiogenesis in Human Umbilical Vein Endothelial Cells

doi: 10.4061/2011/351010

Figure Lengend Snippet: Lysophosphatidic acid- (LPA-) induced lymphatic marker expressions are mediated through endothelial growth factor receptor (EGFR) transactivation-, matrix metalloproteinase- (MMP-), LPA1/3-, and interleukin- (IL-) 1R-dependent pathways in human umbilical vein endothelial cells (HUVECs). Starved HUVECs were pretreated with AG1478 (100 nM), AF12198 (10 nM), GM6001 (10 μ M), Ki16425 (10 μ M), and toxin B (10 nM) for 1 h, followed by LPA (5 μ M) treatment for an additional 8 h. HUVECs were dissociated by trypsinization and fixed with 4% paraformaldehyde. Fixed cells were labeled with antibodies for Prox-1, LYVE-1, and podoplanin, and then analyzed by CyFlow. Cells were also stained with a normal mouse or goat immunoglobulin G antibody, which served as a negative control.

Article Snippet: The Matrigel was fixed, blocked, permeabilized, and stained with rabbit anti-mouse Prox-1 (Abcam), followed by incubation with a goat anti-rabbit AlexaFluor-555-conjugated secondary antibody (Molecular Probes).

Techniques: Marker, Labeling, Staining, Negative Control

Journal: International Journal of Inflammation

Article Title: Interleukin-1 β Expression Is Required for Lysophosphatidic Acid-Induced Lymphangiogenesis in Human Umbilical Vein Endothelial Cells

doi: 10.4061/2011/351010

Figure Lengend Snippet: Lysophosphatidic acid- (LPA-) stimulated endothelial cell tube formation in vitro is mediated through endothelial growth factor receptor (EGFR) transactivation-, matrix metalloproteinase- (MMP-), LPA1/3-, and interleukin- (IL-) 1R-dependent mechanisms. Starved HUVECs were pretreated with AG1478 (100 nM), AF12198 (10 nM), GM6001 (10 μ M), Ki16425 (10 μ M), and toxin B (10 nM) for 1 h, followed by LPA (5 μ M) treatment for an additional 24 h, and then cells were seeded onto Matrigel-coated plates. After 6 h of plating, Matrigel of each experiment was fixed and subjected to an immunocytochemical assay. The Matrigel was stained with PECAM-1 or a Prox-1 primary antibody followed by an FITC-conjugated secondary antibody. Images were taken after plating and visualized by immunofluorescence microscopy.

Article Snippet: The Matrigel was fixed, blocked, permeabilized, and stained with rabbit anti-mouse Prox-1 (Abcam), followed by incubation with a goat anti-rabbit AlexaFluor-555-conjugated secondary antibody (Molecular Probes).

Techniques: In Vitro, Staining, Immunofluorescence, Microscopy

Journal: Scientific Reports

Article Title: New lymphatic cell formation is associated with damaged brain tissue clearance after penetrating traumatic brain injury

doi: 10.1038/s41598-021-89616-3

Figure Lengend Snippet: Immunohistochemical staining images showing PROX1 expression in ( A ) normal mouse brain tissue and ( B – F ) mouse brain tissue on days 3, 7, 15, 21 and 30 days, following pTBI. ( A ) PROX1 is not expressed in normal mouse brain tissue. PROX1 expression (brown; black arrows) was detected around the injury site (red arrow) on days ( B ) 3 and ( C ) 7. ( D ) PROX1-expressing cells and tubular structures (black arrows) were observed around the injury site on day 15 and a small number of black hemosiderin particles (black arrowheads) were visible. ( E , F ) PROX1 expression (black arrows) and black hemosiderin particles (black arrowheads) were still visible at the injury site on days 21 and 30. ( G ) Immunohistochemical results of PROX1 were analyzed by Image-Pro Plus 6.0 software. The average density (IOD) of PROX1-positive cells in normal mouse brain tissue and in mouse brain tissue on days 3, 7, 15, 21 and 30 days, respectively, following pTBI. The n = 10/group was used for calculating the average IOD. Comparisons between groups were performed using the Kruskal–Wallis H test followed by Bonferroni post-hoc analysis.

Article Snippet: The sections were first incubated with rabbit anti-mouse PROX1 antibody (1:100; Boster Biological Technology Co., Wuhan, China), rabbit anti-mouse CD34 antibody (1:100; BosterBiological Technology) or goat anti-mouse LYVE-1 antibody (1:100; R&D Systems, Shanghai, China) at 4 °C for 24 h. Next, the sections were incubated with horseradish peroxidase-labeled goat anti-rabbit IgG or rabbit anti-goat IgG secondary antibodies.

Techniques: Immunohistochemical staining, Staining, Expressing, Software

Journal: Scientific Reports

Article Title: New lymphatic cell formation is associated with damaged brain tissue clearance after penetrating traumatic brain injury

doi: 10.1038/s41598-021-89616-3

Figure Lengend Snippet: Double immunofluorescence staining images showing LYVE-1/PROX1 expression in brain tissue 7 days after pTBI. Expression of ( A ) LYVE-1 (green); ( B ) PROX1 (red); and ( C ) merge (yellow).

Article Snippet: The sections were first incubated with rabbit anti-mouse PROX1 antibody (1:100; Boster Biological Technology Co., Wuhan, China), rabbit anti-mouse CD34 antibody (1:100; BosterBiological Technology) or goat anti-mouse LYVE-1 antibody (1:100; R&D Systems, Shanghai, China) at 4 °C for 24 h. Next, the sections were incubated with horseradish peroxidase-labeled goat anti-rabbit IgG or rabbit anti-goat IgG secondary antibodies.

Techniques: Double Immunofluorescence Staining, Expressing

Journal: IBRO Neuroscience Reports

Article Title: Prospero-related homeobox protein 1 (Prox1) enhances myelin sheath regeneration in injured sciatic nerve

doi: 10.1016/j.ibneur.2025.09.011

Figure Lengend Snippet: Double IF staining of Prox1 (red) and MBP (green) with IF (×200) and their intensity profile obtained using ImageJ software, along an ideal straight line (white).

Article Snippet: Rabbit anti-Myelin basic protein/MBP Polyclonal antibody (Proteintech, USA, catalog # 10458–1-AP); Mouse anti-MBP tag Monoclonal antibody (Proteintech, USA, catalog # 66003–1-Ig); Anti-PROX1 Antibody (Boster Bio, China, catalog # A01985–1); DAB Chromogenic Substrate Reagent Kit (Blue) (Boster Bio, China, catalog # AR1025); Rabbit Anti-GAPDH (Jiangsu KeyGEN BioTECH, China, catalog # KGC6102–1); Luxol Fast Blue Myelin Stain Kit (Solarbio, China, G3240); KeygenMAX 3000 transfection reagent (Jiangsu KeyGEN BioTECH, China, catalog # KGA9705–1); PrimeScriptTM RT Master Mix (TaKaRa, Japan, catalog # RR036B); Olympus Inverted Microscope (OLYMPUS, Japan, catalog # IX51); ABI Stepone real-time PCR (Thermo Fisher, USA); Trans-Blot® TurboTM Transfer System (BIO-RAD, USA, catalog #1704150).

Techniques: Staining, Software

Journal: IBRO Neuroscience Reports

Article Title: Prospero-related homeobox protein 1 (Prox1) enhances myelin sheath regeneration in injured sciatic nerve

doi: 10.1016/j.ibneur.2025.09.011

Figure Lengend Snippet: (A) IHC images of Prox1 proteins in sciatic nerve tissue of control sciatic nerves and injured sciatic nerves on Day 3, 4, 5, 6, 7 and 14 post-injuries (×200) and the quantification of Prox1 proteins in top right panel (P value was normalized to control group, **** P < 0.0001, n = 5, One-Way ANOVA analysis was used). (B) Western blot and quantitative analysis of Prox1 protein expression in sciatic nerve tissue from control nerves and injured nerves at days 3, 4, 5, 6, 7, and 14 post-injury (**** P < 0.0001, n = 3, One-Way ANOVA analysis was used).

Article Snippet: Rabbit anti-Myelin basic protein/MBP Polyclonal antibody (Proteintech, USA, catalog # 10458–1-AP); Mouse anti-MBP tag Monoclonal antibody (Proteintech, USA, catalog # 66003–1-Ig); Anti-PROX1 Antibody (Boster Bio, China, catalog # A01985–1); DAB Chromogenic Substrate Reagent Kit (Blue) (Boster Bio, China, catalog # AR1025); Rabbit Anti-GAPDH (Jiangsu KeyGEN BioTECH, China, catalog # KGC6102–1); Luxol Fast Blue Myelin Stain Kit (Solarbio, China, G3240); KeygenMAX 3000 transfection reagent (Jiangsu KeyGEN BioTECH, China, catalog # KGA9705–1); PrimeScriptTM RT Master Mix (TaKaRa, Japan, catalog # RR036B); Olympus Inverted Microscope (OLYMPUS, Japan, catalog # IX51); ABI Stepone real-time PCR (Thermo Fisher, USA); Trans-Blot® TurboTM Transfer System (BIO-RAD, USA, catalog #1704150).

Techniques: Control, Western Blot, Expressing

Journal: IBRO Neuroscience Reports

Article Title: Prospero-related homeobox protein 1 (Prox1) enhances myelin sheath regeneration in injured sciatic nerve

doi: 10.1016/j.ibneur.2025.09.011

Figure Lengend Snippet: MBP protein level was significantly increased by PROX1 overexpression. (A) qPCR quantification of PROX1 overexpression, (B) WB images of MBP and GAPDH protein from cells transfected with NC, pcDNA-vector or pcDNA-prox1, (C) the quantification of (B). (**** P < 0.0001, n = 3, One-Way ANOVA analysis was used).

Article Snippet: Rabbit anti-Myelin basic protein/MBP Polyclonal antibody (Proteintech, USA, catalog # 10458–1-AP); Mouse anti-MBP tag Monoclonal antibody (Proteintech, USA, catalog # 66003–1-Ig); Anti-PROX1 Antibody (Boster Bio, China, catalog # A01985–1); DAB Chromogenic Substrate Reagent Kit (Blue) (Boster Bio, China, catalog # AR1025); Rabbit Anti-GAPDH (Jiangsu KeyGEN BioTECH, China, catalog # KGC6102–1); Luxol Fast Blue Myelin Stain Kit (Solarbio, China, G3240); KeygenMAX 3000 transfection reagent (Jiangsu KeyGEN BioTECH, China, catalog # KGA9705–1); PrimeScriptTM RT Master Mix (TaKaRa, Japan, catalog # RR036B); Olympus Inverted Microscope (OLYMPUS, Japan, catalog # IX51); ABI Stepone real-time PCR (Thermo Fisher, USA); Trans-Blot® TurboTM Transfer System (BIO-RAD, USA, catalog #1704150).

Techniques: Over Expression, Transfection, Plasmid Preparation