Journal: The Journal of Clinical Investigation

Article Title: TIE2 activation by antibody-clustered endogenous angiopoietin-2 prevents capillary loss and fibrosis in experimental kidney disease

doi: 10.1172/JCI190286

Figure Lengend Snippet: ( A ) Schematic diagram of the experimental setup with UUO, contralateral control kidney (CL), and ABTAA (or IgG) administration. ( B and C ) Schematic diagram of ABTAA function ( B ) and inducible conditional mouse lines used in the study ( C ). TAM, tamoxifen induction. ( D and E ) pTIE2/TIE2 in 3-day UUO kidneys in ABTAA (ABT) and Veptp iECKO (Ve KO ) mice, respectively. Data are based on n = 3–10 mice/group and 3 ( D ) and 1 ( E ) blots (see ). ( F ) VEPTP in kidney lysates from CL and 3-day UUO kidneys in WT and Veptp iECKO mice. Data are based on n = 3 mice/group and 1 blot (see ). ( G ) TIE2 protein measured by ELISA in kidney homogenates from CL and 3-day UUO kidneys in Tie2 iECKO (T2 KO ) mice. Data are based on n = 3–5 mice/group. ( H ) Renal perfusion measured with ultrasound contrast imaging in 2-day UUO/CL kidneys from ABTAA-treated, Veptp iECKO , Tie2 iECKO , and Pdgfb iKO (Pb KO ) mice and their respective controls. The same group of sham-operated mice was used to establish baseline perfusion levels for comparison with UUO-injured kidneys. Data are based on n = 5–8 mice/group. ( I – K ) Immunohistochemistry and quantification from renal cortex for endothelial markers (endomucin [EMCN] and podocalyxin [PODXL]) in 3-day UUO kidneys from ABTAA-treated, Veptp iECKO , Tie2 iECKO , and Pdgfb iKO mice and their controls. Data are based on n = 5–11 mice/group and >600 images/marker. Scale bars: 50 μm. Data represent mean ± SD, and each symbol represents 1 mouse (females, magenta; males, cyan). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001, determined by 1-way ANOVA and Tukey’s post hoc test ( F – H , J , and K ) or unpaired 2-tailed Student’s t test ( D and E ).

Article Snippet: The membranes were stripped and reprobed with goat anti-mouse TIE2 Ab (AF762; R&D Systems).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Imaging, Comparison, Immunohistochemistry, Marker

Journal: The Journal of Clinical Investigation

Article Title: TIE2 activation by antibody-clustered endogenous angiopoietin-2 prevents capillary loss and fibrosis in experimental kidney disease

doi: 10.1172/JCI190286

Figure Lengend Snippet: ( A and B ) Semiquantitative grading of capillary length with fenestrations from micrographs of peritubular capillaries in 3-day UUO kidneys from ABTAA-treated (ABT), Veptp iECKO (Ve KO ), and Tie2 iECKO (T2 KO ) mice. Data are based on n = 4–5 mice/group and scoring of 892 micrographs. Scoring is based on percentage of endothelium with fenestrations as follows: 0, 0%–5%; 1, 6%–25%; 2, 26%–50%; 3, 51%–75%; and 4, 76%–100%. The fenestrated endothelium is indicated by an arrow. Scale bars: 2 μm. ( C ) Quantification of endothelial nuclei in Tie2 iECKO and WT mice from 1-, 3-, and 10-day UUO kidneys. Data are based on n = 4–7 mice/group, and 310 images were quantified. ( D ) Representative image with immunohistochemistry for endomucin (cyan) together with the endothelial Cdh5 -Tdtomato lineage tracer (magenta). Scale bars: 50 μm. Data represent mean ± SD, and each symbol represents 1 mouse (females, magenta; males, cyan). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001, determined by 1-way ANOVA and Tukey’s post hoc test ( B and C ).

Article Snippet: The membranes were stripped and reprobed with goat anti-mouse TIE2 Ab (AF762; R&D Systems).

Techniques: Immunohistochemistry

Journal: The Journal of Clinical Investigation

Article Title: TIE2 activation by antibody-clustered endogenous angiopoietin-2 prevents capillary loss and fibrosis in experimental kidney disease

doi: 10.1172/JCI190286

Figure Lengend Snippet: ( A – C ) Immunohistochemistry and quantifications from renal cortex for fibrosis markers (aSMA and vimentin [VIM]) in 3-day UUO kidneys from ABTAA-treated (ABT), Veptp iECKO (Ve KO ), Tie2 iECKO (T2 KO ), and Pdgfb iKO (Pb KO ) mice and their controls. Data are based on n = 5–11 mice/group and quantifications from >600 images/marker. Scale bars: 50 μm. ( D ) Renal Col1a1 expression in ABTAA-treated mice. Data are based on n = 6 mice/group. ( E ) Renal Pdgfrb expression in Pdgfb iKO mice. Data are based on n = 5–6 mice/group. ( F ) Gene expression of Col1a1 , Tagln , and Fn1 in 3-day UUO kidneys from Tie2 iECKO mice. Data are based in n = 5–6 mice/group. Data represent mean ± SD, and each symbol represents 1 mouse (females, magenta; males, cyan). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001, determined by 1-way ANOVA and Tukey’s post hoc test ( B – F ).

Article Snippet: The membranes were stripped and reprobed with goat anti-mouse TIE2 Ab (AF762; R&D Systems).

Techniques: Immunohistochemistry, Marker, Expressing, Gene Expression

Journal: The Journal of Clinical Investigation

Article Title: TIE2 activation by antibody-clustered endogenous angiopoietin-2 prevents capillary loss and fibrosis in experimental kidney disease

doi: 10.1172/JCI190286

Figure Lengend Snippet: ( A ) Breeding strategy for Tie2 iECKO mice with endothelial lineage tracer and myofibroblast reporter. ( B and C ) Imaging and quantification of Pdgfra -H2BGFP + nuclei (cyan) in renal cortex of 3- and 10-day UUO kidneys in WT and Tie2 iECKO (T2 KO ) mice. Data are based on n = 3–6 mice/group and quantifications from >180 images. Scale bars: 50 μm. ( D ) Representative confocal image stack (30 μm) from renal cortex showing Cdh5 -TdTomato (magenta) and Pdgfra -H2BGFP (cyan) in renal cortex 10 days after UUO in a Tie2 iECKO mouse. Scale bar: 10 μm. ( E ) Confocal images from renal cortex showing tubular epithelial marker NaK/ATPase (blue), Pdgfra -GFP (cyan), and Cdh5 -TdTomato (magenta) 10 days after UUO in WT and Tie2 iECKO mice. Representative images of n = 3 mice per group. Scale bars: 25 μm. Data in graphs represent mean ± SD, and each symbol represents 1 mouse (females, magenta; males, cyan). ** P < 0.01, *** P < 0.001, and **** P < 0.0001, determined by 1-way ANOVA and Tukey’s post hoc test ( C ).

Article Snippet: The membranes were stripped and reprobed with goat anti-mouse TIE2 Ab (AF762; R&D Systems).

Techniques: Imaging, Marker

Journal: The Journal of Clinical Investigation

Article Title: TIE2 activation by antibody-clustered endogenous angiopoietin-2 prevents capillary loss and fibrosis in experimental kidney disease

doi: 10.1172/JCI190286

Figure Lengend Snippet: ( A ) Tubular segments with pathological vacuoles (red arrow) in kidney sections stained with toluidine blue. ( B ) Quantification of tubular segments with vacuoles from ABTAA-treated (ABT), Veptp iECKO (Ve KO ), and Tie2 iECKO (T2 KO ) mice. Data are based on n = 4–5 mice/group and >10,000 tubular cross sections. ( C ) RNA-ISH for Pecam1 (cyan) and Pdgfb (magenta) in 3-day UUO kidneys. Representative image of n = 4 mice. Scale bars: 50 μm. ( D and E ) Gene expression of Kim1 , Pdgfb , Pdgfrb , Tie2 , and Angpt1 in UUO kidneys from indicated time points. Data are based on n = 3–7 mice/group. ( F ) RNA-ISH for Angpt1 (magenta), mesenchymal marker Pdgfrb (cyan), and tubular marker Atp1a1 (blue) in 3-day UUO kidneys. Representative image of n = 3 mice. Scale bars: 50 μm. ( G – I ) Expression of Pdgfb/PDGFB in 3-day UUO kidneys from ABTAA-treated, Veptp iECKO , and Pdgfb iKO (Pb KO ) mice. Data are based on n = 5–9 mice/group. ( J ) Patient data retrieved from Nephroseq for renal CDH5 (endothelial marker), ANGPT2 , PDGFB , and ANGPT1 expression in CKD and renal dysfunction compared with normal human kidney. Data in graphs ( B , D , E , and G – I ) represent mean ± SD, and each symbol represents 1 mouse (females, magenta; males, cyan). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001, determined by 1-way ANOVA. Human data ( J ) represent Log2 expression and statistical differences from Nephroseq (see Methods).

Article Snippet: The membranes were stripped and reprobed with goat anti-mouse TIE2 Ab (AF762; R&D Systems).

Techniques: Staining, Gene Expression, Marker, Expressing

Journal: The Journal of Clinical Investigation

Article Title: TIE2 activation by antibody-clustered endogenous angiopoietin-2 prevents capillary loss and fibrosis in experimental kidney disease

doi: 10.1172/JCI190286

Figure Lengend Snippet: ( A ) Schematic diagram of administration of ABTAA for evaluation in 10-day UUO kidneys. ( B – F ) Immunohistochemistry of renal cortex for capillary density (endomucin [EMCN] and podocalyxin [PODXL]) and tubulointerstitial fibrosis (aSMA and vimentin) in 10-day UUO kidneys from ABTAA-treated mice (ABT) and Tie2 iECKO (T2 KO ) mice. Data are based on n = 6–7 mice/group and quantification of >220 images/marker. Scale bars: 50 μm. ( E ) Protein concentration for PDGFB in 10-day UUO kidneys from ABTAA-treated mice. Data are based on n = 5 mice/group. Data in graphs represent mean ± SD, and each symbol represents 1 mouse (females, magenta; males, cyan). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001, determined by 1-way ANOVA and Tukey’s post hoc test.

Article Snippet: The membranes were stripped and reprobed with goat anti-mouse TIE2 Ab (AF762; R&D Systems).

Techniques: Immunohistochemistry, Marker, Protein Concentration

Journal: Nature cardiovascular research

Article Title: Angiopoietin-TIE2 feedforward circuit promotes PIK3CA-driven venous malformations.

doi: 10.1038/s44161-025-00655-9

Figure Lengend Snippet: Fig. 6 | Increased TIE2 phosphorylation and SMC coverage in Pik3ca-driven VM in mice. a, PLA staining of activated TIE2 on ear skin paraffin sections from Pik3caH1047R;Vegfr1-CreERT2 and Cre− littermate Ctrl mice, detected using pTyr and TIE2 antibodies. DAPI marks cell nuclei. b, Quantification of PLA signals within PECAM1+ blood vessels. Data represent the number of PLA dots per μm2 of PECAM1+ vessel area, mean ± s.d. (n = 8 (Ctrl) and n = 26 (Pik3caH1047R) vessels from four mice per genotype, unpaired two-tailed Student’s t-test, ****P = 0.0000054). c, Immunofluorescence of ear skin paraffin sections from Pik3caH1047R;Vegfr1- CreERT2 and Cre− littermate Ctrl mice using phospho-TIE2 antibodies. d, Phospho- TIE2 signal within EMCN+ vessels, represented as corrected total cell fluorescence (CTFC) of EMCN+ vessel area. Data points represent pTIE2 CTFC, mean ± s.d. (n = 20 (Ctrl) and n = 31 (Pik3caH1047R) vessels from two mice per genotype, unpaired two-tailed Student’s t-test, ****P = 0.000096; Extended Data Fig. 8d). e, Whole-mount immunofluorescence of ear skin from Pik3caH1047R;

Article Snippet: Anti-TIE2 antibody (goat, AF313) was obtained from R&D Systems and used at a dilution of 1:1,000.

Techniques: Phospho-proteomics, Staining, Two Tailed Test, Immunofluorescence, Fluorescence

Journal: Nature cardiovascular research

Article Title: Angiopoietin-TIE2 feedforward circuit promotes PIK3CA-driven venous malformations.

doi: 10.1038/s44161-025-00655-9

Figure Lengend Snippet: Fig. 7 | Increased TIE2 phosphorylation in human PIK3CA-driven VMs. a, H&E-stained paraffin sections of cutaneous VMs from individuals with PIK3CA (top) or TEK (below) mutations. Areas defined as non-lesional (NL) and VM lesions are indicated. b, PLA staining of activated TIE2, detected using pTyr and TIE2 antibodies, in representative vessels from indicated NL and VM regions. DAPI marks cell nuclei. c, Quantification of PLA signals within PECAM1+ veins, represented as mean PLA dots per EC nucleus ± s.d. (n = 7 (PIK3CA), n = 6 (TEK), n = 2 (Ctrl) and n = 5 (NL) individuals, with symbols indicating different mutations; ordinary one-way analysis of variance (ANOVA) and Tukey’s multiple-comparison test, **P(PIK3CA VM versus Ctrl vein) = 0.0027 and **P(TEK VM versus Ctrl vein) = 0.0078; Extended Data Fig. 7). d, Representative immunofluorescence image (left) and quantification (right) showing the

Article Snippet: Anti-TIE2 antibody (goat, AF313) was obtained from R&D Systems and used at a dilution of 1:1,000.

Techniques: Phospho-proteomics, Staining, Comparison, Immunofluorescence

Journal: JOR Spine

Article Title: CD24 Positive Nucleus Pulposus Cells in Adult Human Intervertebral Discs Maintain a More Notochordal Phenotype Than GD2 Positive Cells

doi: 10.1002/jsp2.70029

Figure Lengend Snippet: Antibody details, supplier details and dilutions used for flow cytometry and FACS.

Article Snippet: Secondary antibody incubation was performed using goat anti‐mouse for Tie2 (Goat Anti‐Mouse IgG Antibody (H + L), biotinylated (BA‐9200‐1.5), Vector Laboratories) and goat anti‐rabbit for CD24 and GD2, 1:300 (Goat Anti‐Rabbit IgG (H + L), Biotinylated, BA‐1000‐1.5, Vector Laboratories) for 1 h at RT.

Techniques: Flow Cytometry, Control

Journal: JOR Spine

Article Title: CD24 Positive Nucleus Pulposus Cells in Adult Human Intervertebral Discs Maintain a More Notochordal Phenotype Than GD2 Positive Cells

doi: 10.1002/jsp2.70029

Figure Lengend Snippet: CD24, GD2 and TIE2 immunostaining in a cohort of pediatric and adult NP samples of a range of ages and degeneration grades. Representative images showing immunostaining for CD24 (A), GD2 (C) and TIE2 (E). Scale bars = 100 μm. For each marker, images represent (i) non‐degenerate (grade 0), pediatric NP tissue (10 years old), (ii) mild degeneration (grade 4), adult NP tissue (28 years old); (iii) moderate degeneration (grade 7), elderly NP tissue (73 years old); (iv) severe degeneration (grade 12), elderly NP tissue (65 years old), (v) positive control (CD24: Colon; GD2: Breast carcinoma; TIE2: Testis) and (vi) IgG control (CD24: Colon; GD2: Breast carcinoma; TIE2: Testis). Quantification of marker positivity: (B) CD24, (D) GD2 and (F) TIE2, based on degeneration grade (B, D, F): Pediatric (degeneration grade = 0; n = 6), mild degeneration (2–5; n = 10), moderate degeneration (6–8; n = 10), severe degenerate (9–12; n = 10) and age (A, C, E): Pediatric (10–17, n = 6), young (20–35; n = 10), adult (40–56; n = 10); and elderly (65–75; n = 10). Graphs show mean ± SD. * p < 0.05, one‐way ANOVA, multiple comparisons, test for linear trend, post hoc test.

Article Snippet: Secondary antibody incubation was performed using goat anti‐mouse for Tie2 (Goat Anti‐Mouse IgG Antibody (H + L), biotinylated (BA‐9200‐1.5), Vector Laboratories) and goat anti‐rabbit for CD24 and GD2, 1:300 (Goat Anti‐Rabbit IgG (H + L), Biotinylated, BA‐1000‐1.5, Vector Laboratories) for 1 h at RT.

Techniques: Immunostaining, Marker, Positive Control, Control

Journal: JOR Spine

Article Title: CD24 Positive Nucleus Pulposus Cells in Adult Human Intervertebral Discs Maintain a More Notochordal Phenotype Than GD2 Positive Cells

doi: 10.1002/jsp2.70029

Figure Lengend Snippet: Quantification of CD24, GD2 and TIE2 positive cells in human primary adult NP samples. (A) Bar graph showing mean (±SD) CD24, GD2 and TIE2 positivity in tissue determined using immunostaining ( n = 30) and (B) line graphs comparing immunopositivity in each sample. (C) Bar graph showing mean (±SD) CD24, GD2 and TIE2 positivity in NP cells at passage 1 of culture determined using flow cytometry ( n = 14) and (D) line graphs comparing cell positivity in each sample.

Article Snippet: Secondary antibody incubation was performed using goat anti‐mouse for Tie2 (Goat Anti‐Mouse IgG Antibody (H + L), biotinylated (BA‐9200‐1.5), Vector Laboratories) and goat anti‐rabbit for CD24 and GD2, 1:300 (Goat Anti‐Rabbit IgG (H + L), Biotinylated, BA‐1000‐1.5, Vector Laboratories) for 1 h at RT.

Techniques: Immunostaining, Flow Cytometry

Journal: PLoS ONE

Article Title: Tie2-Expressing Monocytes Are Associated with Identification and Prognoses of Hepatitis B Virus Related Hepatocellular Carcinoma after Resection

doi: 10.1371/journal.pone.0143657

Figure Lengend Snippet: (A) Purified peripheral blood monocytes were stained with conjugated control antibodies (IgG-FITC, IgG-PE) or with CD16-PE antibody alone (B) or with CD14-FITC antibody alone (C) to determine the presence of CD14 + CD16 + monocytes (D). (E) TEMs were distinguished by using IgG-APC control and Tie2-APC antibodies. The percentage of TEMs in peripheral blood CD14 + CD16 + monocytes for this representative sample was 41.2%.

Article Snippet: Then sections were incubated with rabbit anti-human CD14 antibody (sc-9150, Santa Cruz Biotechnology, CA, USA) and goat anti-human Tie2 antibody (sc-31266, Santa Cruz Biotechnology, CA, USA) or mouse anti-human CD16 antibody (sc-20052, Santa Cruz, Santa Cruz Biotechnology, CA, USA).

Techniques: Purification, Staining, Control

Journal: PLoS ONE

Article Title: Tie2-Expressing Monocytes Are Associated with Identification and Prognoses of Hepatitis B Virus Related Hepatocellular Carcinoma after Resection

doi: 10.1371/journal.pone.0143657

Figure Lengend Snippet: (A) Monocytes surface marker CD14 stained with Alexa Fluor 488-labeled IgG (green), Tie2 with Alexa Fluor 594-conjugated IgG (red) and nuclei stained with dapi (blue). Big white arrows represent CD14 + Tie2 + monocytes intratumor (1200×). (B) Monocytes surface marker CD14 stained with Alexa Fluor 488-labeled IgG (green), Tie2 with Alexa Fluor 594-conjugated IgG (red) and nuclei stained with dapi (blue). Thin white arrows represent CD14 + Tie2 + monocytes paratumor (1200×). (C) displayed percentage of TEMs in intratumoral tissue was significantly higher than that in the corresponding paratumoral tissue ( P <0.001).

Article Snippet: Then sections were incubated with rabbit anti-human CD14 antibody (sc-9150, Santa Cruz Biotechnology, CA, USA) and goat anti-human Tie2 antibody (sc-31266, Santa Cruz Biotechnology, CA, USA) or mouse anti-human CD16 antibody (sc-20052, Santa Cruz, Santa Cruz Biotechnology, CA, USA).

Techniques: Marker, Staining, Labeling