Journal: Physiological Reports

Article Title: Establishment of a novel three‐dimensional primary culture model for hippocampal neurogenesis

doi: 10.14814/phy2.13318

Figure Lengend Snippet: Differentiation of neural stem cells into neurons in the Matrigel culture. After seeding the hippocampus fragments in collagen gels under an ALI wells and cultured for 7 days, they were passaged into Matrigel and cultured for 7 days (A). Bright field pictures were taken at day 0 and 7 post‐passage (B). Scale bar: 200 μm. Expression of Nestin (C), DCX (D), a neuronal cell marker, MAP2ab (E), a granule cell marker, Prox‐1 (F) and an astrocytes marker, GFAP (G). Representative immunofluorescence pictures are shown ( n = 4). Scale bar: 50 μm.

Article Snippet: Antibody sources were as follows: Neuronal nuclei (NeuN) (Cell Signaling Technology, Inc., Danvers, MA); Nestin (Immuno‐Biological Laboratories, Gunma, Japan); Doublecortin (DCX) (Sigma‐Aldrich); ΜAP2ab, Prox‐1 (Bioss antibodies, Woburn, MA); Tuj1 (Bethyl Laboratories, Montgomery, TX); Glial Fibrillary Acidic Protein (GFAP) (Diagnostic BioSystems, Pleasanton, CA); Valosin‐containing protein (VCP) (GeneTex, Inc., Irvine, CA).

Techniques: Cell Culture, Expressing, Marker, Immunofluorescence

Journal: Physiological Reports

Article Title: Establishment of a novel three‐dimensional primary culture model for hippocampal neurogenesis

doi: 10.14814/phy2.13318

Figure Lengend Snippet: Expression changes of neuronal differentiation marker proteins in the Matrigel culture. After the hippocampus tissues were cultured in ALI wells and seeded into Matrigel, expression of Nestin (A, n = 4), DCX (B, n = 5), a neuronal cell marker, Tuj1 (C, n = 3) and Prox‐1 (D, n = 4) was determined by Western blotting at day 0 and 7 post‐passaging. Equal protein loading was confirmed using total VCP antibody. * P < 0.05 versus P1 Day 0.

Article Snippet: Antibody sources were as follows: Neuronal nuclei (NeuN) (Cell Signaling Technology, Inc., Danvers, MA); Nestin (Immuno‐Biological Laboratories, Gunma, Japan); Doublecortin (DCX) (Sigma‐Aldrich); ΜAP2ab, Prox‐1 (Bioss antibodies, Woburn, MA); Tuj1 (Bethyl Laboratories, Montgomery, TX); Glial Fibrillary Acidic Protein (GFAP) (Diagnostic BioSystems, Pleasanton, CA); Valosin‐containing protein (VCP) (GeneTex, Inc., Irvine, CA).

Techniques: Expressing, Marker, Cell Culture, Western Blot, Passaging

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