Journal: Journal of cancer research and therapeutics

Article Title: Syk expression in non-small-cell lung cancer and its relation with angiogenesis.

doi: 10.4103/0973-1482.154082

Figure Lengend Snippet: Figure 2: CD34 immunostaining locates in microvascular endothelial cells with brown granular in tumor tissue SABC × 200

Article Snippet: Streptavidin‐biotin complex (SABC) kit, CD34 antibody, and DAB chromogenic reagent were from Wuhan Boster Biotechnology Company.

Techniques: Immunostaining

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 CD34 expression in ( A ) normal mouse brain tissue and ( B – F ) mouse brain tissue on days 3, 7, 15, 21 and 30 days, following the pTBI. ( A ) CD34 was not expressed in normal mouse brain tissue. CD34-positive cells and blood vessels (brown; black arrows) became visible at the injury site (red arrow) on days ( B ) 3 and ( C ) 7. ( D ) After 15 days, CD34 expression at the injury site (red arrow) decreased. ( E ) CD34 expression at the injury site could not be detected on day 21. Instead, black hemosiderin particles (black arrows) were observed at the injury site. ( F ) Some black hemosiderin particles remained visible at the injury site on day 30. ( G ) Immunohistochemical results of CD34 were analyzed by Image-Pro Plus 6.0 software. The average density (IOD) of CD34-positive cells in normal mouse brain tissue and in mouse brain tissue on days 3, 7, 15, 21 and 30 days, 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 CD34/LYVE-1 expression in the brain tissue 3 days after pTBI. Expression of ( A ) LYVE-1 (green); ( B ) CD34 (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: Neural Regeneration Research

Article Title: Radix Ilicis Pubescentis total flavonoids combined with mobilization of bone marrow stem cells to protect against cerebral ischemia/reperfusion injury

doi: 10.4103/1673-5374.177736

Figure Lengend Snippet: RIPTFs elevated the effect of bone marrow stem cell mobilization on Bax, Bcl-2, CD34 and BrdU immunoreactivities in rat brain tissue after cerebral ischemia/reperfusion injury (immunohistochemical staining, × 400). Compared with the model group, Bax immunoreactivity was weaker in nerve cells, but CD34 immunoreactivity noticeably enhanced in the high-dose RIPTFs group. Bcl-2 immunoreactivity was clearly enhanced in the mobilization and high-dose RIPTFs groups. BrdU immunoreactivity was obviously enhanced in the high-, moderate- and low-dose RIPTFs, and mobilization groups. Arrows indicate positive expression. RIPTFs: Radix Ilicis Pubescentis total flavonoids.

Article Snippet: Sections were incubated with rabbit anti-rat BrdU monoclonal antibody (Boster, Wuhan, China), rabbit anti-CD34 polyclonal antibody (1:200) (Boster), rabbit anti-Bax polyclonal antibody (1:200) (Boster), and rabbit anti-Bcl-2 polyclonal antibody (1:200) (Boster) at 4°C overnight.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Cell systems

Article Title: Analysis of Body-wide Unfractionated Tissue Data to Identify a Core Human Endothelial Transcriptome.

doi: 10.1016/j.cels.2016.08.001

Figure Lengend Snippet: Figure 2. Correlation Values between the Reference Endothelial Cell Transcripts CLEC14A, vWF, CD34 and Proteins Des- cribed as EC Enriched in the Literature (A) RNA-seq data from 124 individual samples from 32 different human tissue types were used to generate Spearman pair wise correlation values between the EC reference transcripts CLEC14A, vWF, and CD34 and transcripts reported in the literature as EC enriched. (B) IHC images of salivary gland, gallbladder, and lung tissue stained for proteins encoded by HSPA12B, PECAM1, ENG, ESM1, LIPG, and EDF1. Corresponding scatterplots (right) show the correlation between mean FPKM values and mean EC percentage in selected sequenced tissue samples. Tissue type represented by each symbol corresponds to that indicated on the x axis of Figure 1A. Pearson correlations and correspond- ing p values are shown for each scatterplot. Scale bars, 50 mm.

Article Snippet: Primary antibody against CLEC14A (Atlas Antibodies HPA039468), VWF (Atlas Antibodies HPA001815), CD34 (Atlas Antibodies HPA036722), HSPA12B (Atlas Antibodies HPA013659), ENG (Novocastra NCL-CD105), ESM1 (Atlas Antibodies HPA036660), PTPRC (Atlas Antibodies HPA000440), ITGA2B (Atlas AntibodiesHPA031168),MYH11 (Atlas AntibodiesHPA0145359), EDH2 (AtlasAntibodiesHPA049890), LIMS2 (AtlasAntibodies HPA058340), FAM110D (Atlas Antibodies HPA013664), KANK3 (Atlas Antibodies HPA051153) or GIPC3 (Atlas Antibodies HPA061258) and a dextran polymer visualization system (UltraVision LP HRP polymer, Lab Vision) were incubated for 30 min each at room temperature and slides were developed for 10 min using Diaminobenzidine (Lab Vision) as the chromogen.

Techniques: RNA Sequencing, Staining

Journal:

Article Title: Bone marrow derivation of pericryptal myofibroblasts in the mouse and human small intestine and colon

doi:

Figure Lengend Snippet: Y chromosome detection in the mouse. The Y chromosome is seen as a punctate brown/black density in the cell nucleus. Red staining of the cell cytoplasm identifies positive immunohistochemical staining. (A) Male positive control showing Y chromosome detection in colonic pericryptal myofibroblast cells immunostained with the α smooth muscle actin (αSMA) antibody. (B) Female control demonstrating lack of Y chromosome detection within colonic pericryptal myofibroblasts immunostained with the αSMA antibody. (C–G) Female mouse after male whole bone marrow transplantation showing Y chromosome positive myofibroblasts immunostained with the αSMA antibody. (C) Few αSMA immunoreactive Y chromosome positive cells present in the mouse colon seven days after male bone marrow transplant. (D) Numerous Y chromosome positive myofibroblasts (for example, arrows, see also inserts for high power views) immunoreactive for αSMA in female mouse colon 14 days after male bone marrow transplant. (E) Y chromosome positive myofibroblasts stained with αSMA in the colon at six weeks after transplant showing extension of the column right up to the top of the crypts. (F, G) High power view of Y chromosome positive myofibroblasts that are αSMA immunoreactive in female mouse colon six weeks after transplant. (H) Desmin positive cells within the lamina propria were not Y chromosome positive. (I) Y chromosome positive cells that are negative for the haematopoeitic progenitor marker CD34 in the mouse colon six weeks after transplant. (J) Y chromosome positive cells that are negative for the mouse macrophage marker F4/80 antigen (positively stained macrophages shown by arrows).

Article Snippet: As further tests of the phenotype of bone marrow graft derived cells, we combined Y chromosome in situ hybridisation and immunohistochemistry for desmin (monoclonal antibody MDEII; Euro-Diagnostica, Arnhem, the Netherlands), a murine macrophage marker (rat polyclonal anti-mouse F4/80 antigen; MCAP497; Serotec, Kedlington, Oxford, UK) and the haematopoietic progenitor marker CD34 (rat polyclonal anti-mouse CD34; CL8927 AP; Cedarlane, Ontario, Canada).

Techniques: Staining, Immunohistochemical staining, Positive Control, Transplantation Assay, Marker

Journal: Translational Cancer Research

Article Title: The suppressing effects of VEGF-mediated angiogenesis at different administration sequences of apatinib and transarterial embolization in vivo

doi: 10.21037/tcr.2019.12.97

Figure Lengend Snippet: Immunohistochemical staining showed a positive CD34 reaction (×200). Positive CD34 was identified by dark-brown staining. (A) Control group (CG); (B) combined Group 1 (CG1); (C) combined Group 2 (CG2); (D) combined Group 3 (CG3).

Article Snippet: Both rabbit anti-rat VEGF-165 polyclonal antibody and rabbit anti-rat CD34 polyclonal antibody were purchased from BOSTER, Inc, Wuhan, China.

Techniques: Immunohistochemical staining, Staining, Control

Journal: Cells

Article Title: The Role of Heparan Sulfate and Neuropilin 2 in VEGFA Signaling in Human Endothelial Tip Cells and Non-Tip Cells during Angiogenesis In Vitro

doi: 10.3390/cells10040926

Figure Lengend Snippet: Effect of SULF2 knockdown on VEGFA121- and VEGFA165-induced tip cell formation and sprouting. ( a ) Microarray analysis showed enrichment of SULF2 mRNA in CD34 + cells as compared to CD34 - cells. HUVECs were treated with siNT, si SULF2 or left untreated (Control) and were subsequently cultured in the presence or absence of VEGFA 165 or VEGF 121 . ( b ) Treatment of cells with siSULF2 resulted in a significant reduction in mRNA levels of 87% as compared to treatment with siNT. ( c ) Flow cytometric analysis of CD34 + cells (% of total number of cells). Average number ( d ) and length in µm ( e ) of sprouts per HUVEC spheroid. ( f ) Representative images of sprout formation from HUVEC spheroids. Results are presented as the mean ± standard deviation of three HUVEC donors. Unpaired T-tests ( a , b ), two-way ANOVA ( c ) or two-way mixed ANOVA ( d , e ) were used to calculate statistical differences (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). Scale bar = 250 µm.

Article Snippet: Next, cells were incubated with anti-VEGFR2 (AF357, 1:100) or anti-NRP2 (AF2215, 1:100), both from R&D Systems, anti-VE-cadherin (361900, Invitrogen, 1:100), and anti-CD34 (Sanquin, 1:100) diluted in primary antibody diluent (APG500, Scytek, Logan, UT, USA) for 1 h at room temperature.

Techniques: Microarray, Cell Culture, Standard Deviation

Journal: Cells

Article Title: The Role of Heparan Sulfate and Neuropilin 2 in VEGFA Signaling in Human Endothelial Tip Cells and Non-Tip Cells during Angiogenesis In Vitro

doi: 10.3390/cells10040926

Figure Lengend Snippet: Effects of SULF2 knockdown on VEGFA 121 - and VEGFA 165 -induced cell proliferation and apoptosis. HUVECs were cultured in the presence or absence of VEGFA 165 , VEGF 121 or no VEGF (Control) after treatment with siNT or si SULF2 . Flow cytometric analysis determined the percentage of cells in G0/G1 phase ( a ), S-phase ( b ), and G2/M phase ( c ). FACS analysis after annexin-5 and CD34 labeling determined the percentage of CD34 - non-tip cells ( d ) and CD34 + tip cells ( e ). Results are presented as the mean ± standard deviation of three HUVEC donors. Two-way ANOVA was used to calculate statistical differences (* p < 0.05, *** p < 0.001).

Article Snippet: Next, cells were incubated with anti-VEGFR2 (AF357, 1:100) or anti-NRP2 (AF2215, 1:100), both from R&D Systems, anti-VE-cadherin (361900, Invitrogen, 1:100), and anti-CD34 (Sanquin, 1:100) diluted in primary antibody diluent (APG500, Scytek, Logan, UT, USA) for 1 h at room temperature.

Techniques: Cell Culture, Labeling, Standard Deviation

Journal: Cells

Article Title: The Role of Heparan Sulfate and Neuropilin 2 in VEGFA Signaling in Human Endothelial Tip Cells and Non-Tip Cells during Angiogenesis In Vitro

doi: 10.3390/cells10040926

Figure Lengend Snippet: Effect of NRP2 knockdown on VEGF-induced tip cell formation and sprouting. ( a ) Microarray analysis showed enrichment of NRP2 mRNA in CD34 + cells as compared to CD34 - cells. HUVECs were cultured in the presence or absence of VEGFA 121 and/or VEGFA 165 or after treatment with siNT or si NRP2 ; ( b ) Treatment of cells with si NRP2 resulted in a significant reduction in mRNA levels of 84% as compared to treatment with siNT; ( c ) Treatment of cells with si NRP2 resulted in a significant reduction in protein levels of 60% as compared to treatment with siNT. A representative western blot is shown. Quantification of NRP2 levels as compared to β-actin levels is shown below in three HUVEC donors; ( d ) Flow cytometric analysis of CD34 + cells (% of total number of cells). Average number ( e ) and length in µm ( f ) of sprouts per HUVEC spheroid; ( g ) Representative images of sprout formation from HUVEC spheroids. Results are presented as the mean ± standard deviation of three HUVEC donors. Unpaired T-tests ( a , b ) or Two-way ANOVA analyses ( c – e ) were used to calculate statistical differences (* p < 0.05, ** p < 0.01, **** p < 0.0001). Scale bar = 250 µm.

Article Snippet: Next, cells were incubated with anti-VEGFR2 (AF357, 1:100) or anti-NRP2 (AF2215, 1:100), both from R&D Systems, anti-VE-cadherin (361900, Invitrogen, 1:100), and anti-CD34 (Sanquin, 1:100) diluted in primary antibody diluent (APG500, Scytek, Logan, UT, USA) for 1 h at room temperature.

Techniques: Microarray, Cell Culture, Western Blot, Standard Deviation

Journal: Cells

Article Title: The Role of Heparan Sulfate and Neuropilin 2 in VEGFA Signaling in Human Endothelial Tip Cells and Non-Tip Cells during Angiogenesis In Vitro

doi: 10.3390/cells10040926

Figure Lengend Snippet: Changes in protein levels of NRP2 and VEGFR2 after SULF2 knockdown. HUVECs were transfected with si SULF2 or siNT as a control, seeded on coverslips and cultured for three days. After 2% PFA fixation, cells were stained with antibodies against ( a ) NRP2 or ( b ) VEGFR2 (red), CD34 (green), or VEcad (cyan) and with DAPI (blue). Representative images are shown for experiments on HUVECs of three donors. A 2.5-fold magnification of the area in the red box is presented. Scale bar = 50 µm. Quantification of NRP2 ( c ) and VEGFR2 ( d ) is shown below, relative to siNT with mean ± SD. ** p < 0.01, **** p < 0.0001.

Article Snippet: Next, cells were incubated with anti-VEGFR2 (AF357, 1:100) or anti-NRP2 (AF2215, 1:100), both from R&D Systems, anti-VE-cadherin (361900, Invitrogen, 1:100), and anti-CD34 (Sanquin, 1:100) diluted in primary antibody diluent (APG500, Scytek, Logan, UT, USA) for 1 h at room temperature.

Techniques: Transfection, Cell Culture, Staining

Journal: Cells

Article Title: The Role of Heparan Sulfate and Neuropilin 2 in VEGFA Signaling in Human Endothelial Tip Cells and Non-Tip Cells during Angiogenesis In Vitro

doi: 10.3390/cells10040926

Figure Lengend Snippet: Knockdown of SULF2 leads to increased uptake of VLDL in non-tip cells. ( a ) Demonstration of endosomal uptake of fluorescently-labeled VLDL by CD34 - but not CD34 + cells. Staining of CD34 (tip cells, green), VLDL (cyan), endosomes (EEA, magenta), and nuclei (DAPI, blue) in HUVECs and overlaid images at low (bar = 50 µm) and higher (bar = 20 µm) magnification. ( b – d ) Quantification of total, endosomal, and non-endosomal uptake of fluorescently-labeled VLDL by HUVECs after siRNA-mediated knockdown of SULF2 . Heparanase was used as a control as it removes all HS chains from HSPG proteins. Cells were stained with DAPI and EEA1 for nuclei and endosomes. The numbers of VLDL particles per cell were counted using Matlab and overlap with EEA1 staining was considered to indicate endosomal uptake. Students ’ t -tests were used to calculate statistical differences (* p < 0.05, ** p < 0.01, *** p < 0.001) using HUVECs of three donors and at least seven cells per condition in each experiment.

Article Snippet: Next, cells were incubated with anti-VEGFR2 (AF357, 1:100) or anti-NRP2 (AF2215, 1:100), both from R&D Systems, anti-VE-cadherin (361900, Invitrogen, 1:100), and anti-CD34 (Sanquin, 1:100) diluted in primary antibody diluent (APG500, Scytek, Logan, UT, USA) for 1 h at room temperature.

Techniques: Labeling, Staining

Journal: Cells

Article Title: The Role of Heparan Sulfate and Neuropilin 2 in VEGFA Signaling in Human Endothelial Tip Cells and Non-Tip Cells during Angiogenesis In Vitro

doi: 10.3390/cells10040926

Figure Lengend Snippet: Effect of VLDL on tip cell formation, sprouting, and apoptosis. HUVECs were cultured in the presence or absence of VEGFA 165 and/or VLDL or after treatment with siNT or si SULF2 . ( a ) Flow cytometric analysis of CD34 + cells (% of total number of cells). Average number ( b ) and length in µm ( c ) of sprouts per HUVEC spheroid. Relative amount of apoptotic CD34 - ( d ) and CD34 + cells ( e ). Results are presented as the mean ± standard deviation of three HUVEC donors. Two-way ANOVA was used to calculate statistical differences (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Next, cells were incubated with anti-VEGFR2 (AF357, 1:100) or anti-NRP2 (AF2215, 1:100), both from R&D Systems, anti-VE-cadherin (361900, Invitrogen, 1:100), and anti-CD34 (Sanquin, 1:100) diluted in primary antibody diluent (APG500, Scytek, Logan, UT, USA) for 1 h at room temperature.

Techniques: Cell Culture, Standard Deviation