Journal: Signal Transduction and Targeted Therapy

Article Title: An aptamer-drug conjugate for promising cancer therapy with comprehensive evaluation from rodents to non-human primates

doi: 10.1038/s41392-025-02399-1

Figure Lengend Snippet: Characterization of Sgc8c-M in synthesis, targeting, internalization, and cytotoxicity in vitro. a Chemical structure, HPLC, and mass spectrum of Sgc8c-M. b Surface plasmon resonance (SPR) analysis of Sgc8c-M for binding to recombinant PTK7 proteins from humans, mice, rats, and cynomolgus monkeys at 400 nM. c Flow cytometry analysis of Cy5-labeled Ctrl, Sgc8c, Ctrl-M, and Sgc8c-M (250 nM) for binding to PTK7-positive SUM159 and MDA-MB-468 cells, as well as PTK7-negative Ramos cells. d Flow cytometry analysis of the internalization and endocytic pathway of Cy5-Sgc8c-M in SUM159 cells. The endocytic pathway was determined by measuring the uptake upon the application of various inhibitors. M-β-CD (0.2 mM), CPZ (25 μM), and AMI (100 μM) were preincubated with cells for 30 min at 37 °C; then 400 nM Cy5-Sgc8c-M were added and incubation continued for another 2 h ( n = 3). Unpaired t test was used in comparison to control, ** P < 0.01, *** P < 0.001, ns, not significant. e Confocal microscopy analysis of Cy5-Sgc8c-M (400 nM) for internalization in SUM159 cells at 0.5 h and 2 h. Scale bars, 10 μm. f Cytotoxicity of VcMMAE and Sgc8c-M in SUM159 and MDA-MB-468 cells after a 72-h incubation ( n = 3). Data are presented as mean ± SEM

Article Snippet: The primary antibody, anti-PTK7 rabbit polyclonal antibody (#A9839, abclonal), was diluted to a concentration of 1:250 and applied to the sections for overnight incubation at 4 °C.

Techniques: In Vitro, SPR Assay, Binding Assay, Recombinant, Flow Cytometry, Labeling, Incubation, Comparison, Control, Confocal Microscopy

Journal: Signal Transduction and Targeted Therapy

Article Title: An aptamer-drug conjugate for promising cancer therapy with comprehensive evaluation from rodents to non-human primates

doi: 10.1038/s41392-025-02399-1

Figure Lengend Snippet: In vivo antitumor effect of Sgc8c-M in TNBC. a IHC characterization of PTK7 expression on tumor tissues of SUM159, MDA-MB-468, and TNBC PDX models. Scale bars, 20 μm. b Tumor volume and body weight of SUM159 tumor-bearing mice from a dose screening trial of Sgc8c-M ( n = 4, one-way ANOVA). c , d , f Tumor growth curves for SUM159 ( n = 4), MDA-MB-468 ( n = 4, unpaired t test), and TNBC PDX ( n = 5, unpaired t test) tumor-bearing mice after multiple dosing of 7 mg/kg Sgc8c-M (intravenous injection, i.v.), 10 mg/kg paclitaxel (intraperitoneal injection, i.p.), or 0.359 mg/kg MMAE (i.v.). e , g Tumor-killing effect study of Sgc8c-M in MDA-MB-468 ( n = 3, unpaired t test) and TNBC PDX ( n = 3, unpaired t test). Mice were given 7 mg/kg Sgc8c-M intravenously from day 0, and immunohistochemical characterization of CK19, Ki67, and pHH3 was performed at 48–96 h (48 h for MDA-MB-468, 96 h for TNBC PDX) at the end of administration. Scale bars, 20 μm. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01

Article Snippet: The primary antibody, anti-PTK7 rabbit polyclonal antibody (#A9839, abclonal), was diluted to a concentration of 1:250 and applied to the sections for overnight incubation at 4 °C.

Techniques: In Vivo, Expressing, Injection, Immunohistochemical staining

Journal: Signal Transduction and Targeted Therapy

Article Title: An aptamer-drug conjugate for promising cancer therapy with comprehensive evaluation from rodents to non-human primates

doi: 10.1038/s41392-025-02399-1

Figure Lengend Snippet: Antitumor effect of Sgc8c-M in pancreatic cancer. a Cytotoxicity of VcMMAE, Sgc8c-M, and MMAE in MIA PaCa-2 cells after a 72-h incubation ( n = 3). b , e , f Antitumor effects of Sgc8c-M for MIA PaCa-2, PDX F4, and PDX F5 in vivo ( n = 5, unpaired t test). Tumor-bearing mice were dosed by i.v. every four days for 5–6 cycles with 7 mg/kg Sgc8c-M or 0.359 mg/kg MMAE. PTK7 IHC characterization and hematoxylin-eosin (HE) staining on these tumors showed the differences in PTK7 expression and tissue structure. Scale bars, 20 μm. c MMAE concentration in MIA PaCa-2 tumors at 2 h after a single dose of 7 mg/kg Sgc8c-M or 0.359 mg/kg MMAE ( n = 3, unpaired t test). d Tumor-killing effect study of Sgc8c-M and MMAE in MIA PaCa-2 xenografts. Mice received 7 mg/kg Sgc8c-M or 0.359 mg/kg MMAE every four days for three cycles (q4d × 3) with Ki67 and pHH3 analyzed 24 h after the final dose ( n = 3, one-way ANOVA). Scale bars, 20 μm. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01

Article Snippet: The primary antibody, anti-PTK7 rabbit polyclonal antibody (#A9839, abclonal), was diluted to a concentration of 1:250 and applied to the sections for overnight incubation at 4 °C.

Techniques: Incubation, In Vivo, Staining, Expressing, Concentration Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: An aptamer-drug conjugate for promising cancer therapy with comprehensive evaluation from rodents to non-human primates

doi: 10.1038/s41392-025-02399-1

Figure Lengend Snippet: Antitumor activity of Sgc8c-M in HT-29, NCI-H1975, and OVCAR3 xenograft models. In vivo therapeutic efficacy of Sgc8c-M in HT-29 ( n = 4, one-way ANOVA) ( a ), NCI-H1975 ( n = 3) ( b ), and OVCAR3 ( n = 3, one-way ANOVA) ( c ) xenografts. Tumor-bearing mice were dosed with 7 mg/kg Sgc8c-M (i.v.) or 10 mg/kg paclitaxel (i.p.) when the tumor size reached an average of 100–200 mm 3 . Histology and PTK7 expression in these models were shown as HE staining and IHC staining on tumor tissues of untreated mice, respectively. d The antitumor effects of Sgc8c-M (3.5 mg/kg) and PTK7-targeted ADC h6M24-VcMMAE (DAR4, 5 mg/kg) in the OVCAR3 model ( n = 5, unpaired t test). Scale bars, 20 μm. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01

Article Snippet: The primary antibody, anti-PTK7 rabbit polyclonal antibody (#A9839, abclonal), was diluted to a concentration of 1:250 and applied to the sections for overnight incubation at 4 °C.

Techniques: Activity Assay, In Vivo, Drug discovery, Expressing, Staining, Immunohistochemistry

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Bioinformatics Analysis Identifies Protein Tyrosine Kinase 7 (PTK7) as a Potential Prognostic and Therapeutic Biomarker in Stages I to IV Hepatocellular Carcinoma

doi: 10.12659/MSM.917142

Figure Lengend Snippet: Effect of PTK7 expression in hepatocellular carcinoma (HCC). ( A ) The effect of commonly deregulated HCC target genes on patient overall survival (OS). ( B ) The expression of PTK7 in HCC tumor tissue and adjacent normal tissue samples in cancer at different stages. ( C ) Kaplan-Meier survival curve of PTK7 expression in patients with HCC. ( D ) The receiver operating characteristic (ROC) curve analysis of PTK7 gene expression between HCC and adjacent normal samples. The area under the curve (AUC) that approximates to 1 indicates a greater difference between the tumor and adjacent samples. ( E ) Quantitative reverse transcription PCR (RT-qPCR) of PTK7 in normal liver cells (HL-7702) and HCC cell lines (HepaRG, HepG2, SK-Hep1, and Huh7). ( F ) Western blot of PTK7 in normal liver cells (HL-7702) and HCC cell lines (HepaRG, HepG2, SK-Hep1, and Huh7).

Article Snippet: After blocking with dried skimmed milk powder, the blots were probed with a mouse antibody to alpha-tubulin (Proteintech, Manchester, UK) and a rabbit antibody to human protein tyrosine kinase 7 (PTK7) (Proteintech, Manchester, UK).

Techniques: Expressing, Gene Expression, Reverse Transcription, Quantitative RT-PCR, Western Blot

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Bioinformatics Analysis Identifies Protein Tyrosine Kinase 7 (PTK7) as a Potential Prognostic and Therapeutic Biomarker in Stages I to IV Hepatocellular Carcinoma

doi: 10.12659/MSM.917142

Figure Lengend Snippet: Univariate Cox proportional hazards regression analysis of PTK7, clinicopathological parameters, and overall survival (OS) in patients with hepatocellular carcinoma (HCC).

Article Snippet: After blocking with dried skimmed milk powder, the blots were probed with a mouse antibody to alpha-tubulin (Proteintech, Manchester, UK) and a rabbit antibody to human protein tyrosine kinase 7 (PTK7) (Proteintech, Manchester, UK).

Techniques:

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Bioinformatics Analysis Identifies Protein Tyrosine Kinase 7 (PTK7) as a Potential Prognostic and Therapeutic Biomarker in Stages I to IV Hepatocellular Carcinoma

doi: 10.12659/MSM.917142

Figure Lengend Snippet: Multivariate Cox proportional hazards regression of PTK7 on overall survival (OS).

Article Snippet: After blocking with dried skimmed milk powder, the blots were probed with a mouse antibody to alpha-tubulin (Proteintech, Manchester, UK) and a rabbit antibody to human protein tyrosine kinase 7 (PTK7) (Proteintech, Manchester, UK).

Techniques:

Journal: Cell Death & Disease

Article Title: FOXP4-mediated induction of PTK7 activates the Wnt/β-catenin pathway and promotes ovarian cancer development

doi: 10.1038/s41419-024-06713-7

Figure Lengend Snippet: A The mRNA correlation between FOXP4 and PTK7 in OV from TCGA database. B The spearman correlation between the expression of FOXP4 and PTK7 in all tumor types in TCGA. Red represents positive expression and green represents negative expression. C Real-time PCR analysis of FOXP4 and PTK7 mRNA levels in A2780 cells transfected with EV or FOXP4 expression vector, n = 3. ** p < 0.01 vs EV, *** p < 0.001 vs EV. D Immunoblot analysis showing FOXP4 and PTK7 protein expression in A2780 cells transfected with EV or FOXP4. E Immunoblot analysis of FOXP4 in A2780 and OVCAR8 cells stably expressing either pLKO or shRNAs targeting FOXP4. F Schematic diam shows human PTK7 gene promoter and a putative FOXP4 binding site. TSS: transcription start site. G Schematic diam shows the human PTK7 WT and Mut promoters. The potential binding sites for FOXP4 were highlighted with yellow. H The activities of the human PTK7 WT and Mut promoters in 293T cells transfected with EV or FOXP4 was measured, n = 3. *** p < 0.001 vs EV. I Two shFOXP4s were co-transfected with the indicated plasmid into 293T cells for 36 h. The luciferase activity was then measured, n = 3. *** p < 0.001 vs pLKO. J , K Chip-PCR assay detecting FOXP4 binding to the human PTK7 promoter in A2780 cells expressing stable FOXP4, n = 3. *** p < 0.001 vs IgG. L A2780 cells were treated with ICG-001 or LiCl to detect PTK7 protein levels. M The luciferase activities of TCF/LEF1 in A2780 and OVCAR8 cells transfected with pLKO or shRNAs-PTK7 were measured, n = 3. ** p < 0.01 vs pLKO, *** p < 0.001 vs pLKO. N The luciferase activities of TCF/LEF1 in A2780 and OVCAR8 cells transfected with EV or PTK7 were measured, n = 3. ** p < 0.01 vs EV.

Article Snippet: The following primary antibodies were used: anti-β-catenin (Santa Cruz, sc-7963), anti-FOXP4 (Santa Cruz, sc-390892), anti-PTK7 (Bethyl Laboratories, A304-451A), anti-Cyclin D1 (Abcam, ab134175), anti-GSK3B (Abcam, ab76225), anti-c-Myc (Abcam, ab32072), anti-E-cadherin (Abcam, ab40772), anti-N-cadherin (Abcam, AB207608), anti-Vimentin (Abcam, ab92547), and anti-β-actin (Santa Cruz, sc-8432).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation, Western Blot, Stable Transfection, Binding Assay, Luciferase, Activity Assay

Journal: Cell Death & Disease

Article Title: FOXP4-mediated induction of PTK7 activates the Wnt/β-catenin pathway and promotes ovarian cancer development

doi: 10.1038/s41419-024-06713-7

Figure Lengend Snippet: A PTK7 was retrovirally infected into A2780 cells stably expressing FOXP4-shRNA. Lysates of A2780 cells with pLKO, sh-FOXP4, and sh-FOXP4 + PTK7 were immunoblotted with the indicated antibodies. B The cell growth curve of cells from ( A ), n = 3. ** p < 0.01. C Incorporation of BrdU in cells from ( A ) was measured by ELISA, n = 3. ** p < 0.01, *** p < 0.001. D Clonogenic assay of cells in ( A ), n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001. E Relative migration rates of cells in ( A ) in a transwell assay, n = 3. ** p < 0.01, *** p < 0.001. F A2780 cells stably expressing pLKO or sh-FOXP4 and stably expressing sh-FOXP4 + PTK7 were inoculated subcutaneously in nude mice, respectively, and the folded line graphs indicate the average volume of tumors over time, n = 6. *** p < 0.001. G Image representing the tumor on day 28, n = 6. H Bar graphs represent the weight of the tumor on day 28, n = 6. *** p < 0.001.

Article Snippet: The following primary antibodies were used: anti-β-catenin (Santa Cruz, sc-7963), anti-FOXP4 (Santa Cruz, sc-390892), anti-PTK7 (Bethyl Laboratories, A304-451A), anti-Cyclin D1 (Abcam, ab134175), anti-GSK3B (Abcam, ab76225), anti-c-Myc (Abcam, ab32072), anti-E-cadherin (Abcam, ab40772), anti-N-cadherin (Abcam, AB207608), anti-Vimentin (Abcam, ab92547), and anti-β-actin (Santa Cruz, sc-8432).

Techniques: Infection, Stable Transfection, Expressing, shRNA, Enzyme-linked Immunosorbent Assay, Clonogenic Assay, Migration, Transwell Assay

Journal: Cell Death & Disease

Article Title: FOXP4-mediated induction of PTK7 activates the Wnt/β-catenin pathway and promotes ovarian cancer development

doi: 10.1038/s41419-024-06713-7

Figure Lengend Snippet: A The FOXP4 retrovirus was infected into A2780 cells stably expressing sh-PTK7, and A2780 cell lysates of pLKO, FOXP4, and FOXP4+sh-PTK7 were immunobloted with the indicated antibodies. B The cell growth curve of cells from ( A ), n = 3. *** p < 0.001. C Incorporation of BrdU in cells from ( A ) was measured by ELISA, n = 3. * p < 0.05, ** p < 0.01. D Clonogenic assay of cells in ( A ), n = 3. *** p < 0.001. E Relative invasion rates of cells in ( A ) in a transwell assay, n = 3. *** p < 0.001. F A2780 cells stably expressing EV or FOXP4 or FOXP4 infected with sh-PTK7 in FOXP4 were inoculated subcutaneously in nude mice, respectively, and the folded line graphs indicate the average volume of tumors over time, n = 6. *** p < 0.001. G Image representing the tumor on day 28, n = 6. H Bar graphs represent the weight of the tumor on day 28, n = 6. *** p < 0.001.

Article Snippet: The following primary antibodies were used: anti-β-catenin (Santa Cruz, sc-7963), anti-FOXP4 (Santa Cruz, sc-390892), anti-PTK7 (Bethyl Laboratories, A304-451A), anti-Cyclin D1 (Abcam, ab134175), anti-GSK3B (Abcam, ab76225), anti-c-Myc (Abcam, ab32072), anti-E-cadherin (Abcam, ab40772), anti-N-cadherin (Abcam, AB207608), anti-Vimentin (Abcam, ab92547), and anti-β-actin (Santa Cruz, sc-8432).

Techniques: Infection, Stable Transfection, Expressing, Enzyme-linked Immunosorbent Assay, Clonogenic Assay, Transwell Assay

Journal: Cell Death & Disease

Article Title: FOXP4-mediated induction of PTK7 activates the Wnt/β-catenin pathway and promotes ovarian cancer development

doi: 10.1038/s41419-024-06713-7

Figure Lengend Snippet: In this working model, we depict a signaling pathway involving Wnt/β-catenin/TCF4-FOXP4-PTK7. FOXP4 acts as a transcriptional target of Wnt/β-catenin/TCF4 signaling and enhances the transcription of PTK7, consequently promoting the development and progression of OV tumors.

Article Snippet: The following primary antibodies were used: anti-β-catenin (Santa Cruz, sc-7963), anti-FOXP4 (Santa Cruz, sc-390892), anti-PTK7 (Bethyl Laboratories, A304-451A), anti-Cyclin D1 (Abcam, ab134175), anti-GSK3B (Abcam, ab76225), anti-c-Myc (Abcam, ab32072), anti-E-cadherin (Abcam, ab40772), anti-N-cadherin (Abcam, AB207608), anti-Vimentin (Abcam, ab92547), and anti-β-actin (Santa Cruz, sc-8432).

Techniques:

Journal: Biomaterials Research

Article Title: Targeting treatment of bladder cancer using PTK7 aptamer-gemcitabine conjugate

doi: 10.1186/s40824-022-00328-9

Figure Lengend Snippet: Schematic diagram and application of PTK7-GEMs. A Schematic diagram showing GEM is transformed into a modular drug-type nucleotide and then combined with PTK7-Aptamer to produce PTK7-GEMs. B Application and anti-tumor effect of PTK7-GEMs in three different tumor models

Article Snippet: Thereafter, the membranes were reacted with rabbit polyclonal antibody recognizing PTK7 (Abcam; 1:500) or ACTB (quality control, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:800) overnight at 4 °C.

Techniques: Transformation Assay

Journal: Biomaterials Research

Article Title: Targeting treatment of bladder cancer using PTK7 aptamer-gemcitabine conjugate

doi: 10.1186/s40824-022-00328-9

Figure Lengend Snippet: Serum stability and cell binding ability of PTK7-GEMs. Serum stability analysis of PTK7-GEMs using 12% polyacrylamide gel electrophoresis (PAGE). A PTK7-GEMs were incubated for 0, 1, 2, 4, 8, 12, 16, and 24 h 10% FBS-supplemented or 10% NMS-supplemented RPMI-1640 medium, respectively. B The abscissa indicates that the same sample of PTK7-GEMs was incubated in 10% NMS (red) and 10% FBS (blue) for 0, 1, 2, 4, 8, 12, 16, and 24 h respectively, and the ordinate indicates the respective degradation percentage of the samples. The binding ability of PTK7-GEMs to PTK7 overexpressing cells at 4 °C, as determined using flow cytometry. C , D The binding ability of various FITC-labeled samples to 5637 cells and SV-HUC-1 cells

Article Snippet: Thereafter, the membranes were reacted with rabbit polyclonal antibody recognizing PTK7 (Abcam; 1:500) or ACTB (quality control, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:800) overnight at 4 °C.

Techniques: Binding Assay, Polyacrylamide Gel Electrophoresis, Incubation, Flow Cytometry, Labeling

Journal: Biomaterials Research

Article Title: Targeting treatment of bladder cancer using PTK7 aptamer-gemcitabine conjugate

doi: 10.1186/s40824-022-00328-9

Figure Lengend Snippet: CCK-8 assays of PTK7-GEMs cytotoxicity. A , B Viability of 5637 cells and SV-HUC-1 cells respectively treated with LIB-GEMs, GEM or PTK7-GEMs evaluated by CCK8 assay. C IC 50 of PTK7-GEMs and GEM against 5637 cells (NS, not significant). Data represents the mean ± SEM, n = 3. D , E Cell cycle distributions of 5637 and SV-HUC-1 cells treated with LIB-GEMs, GEM or PTK7-GEMs respectively for 8 h and then incubated for 72 h with complete medium before being tested using flow cytometry

Article Snippet: Thereafter, the membranes were reacted with rabbit polyclonal antibody recognizing PTK7 (Abcam; 1:500) or ACTB (quality control, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:800) overnight at 4 °C.

Techniques: CCK-8 Assay, Incubation, Flow Cytometry

Journal: Biomaterials Research

Article Title: Targeting treatment of bladder cancer using PTK7 aptamer-gemcitabine conjugate

doi: 10.1186/s40824-022-00328-9

Figure Lengend Snippet: Investigation of the GEM release mechanisms from PTK7-GEMs. The profile of PTK7-GEMs release from A Crude cell lysate (untreated by inhibitors) or B Lysate of cells treated with phosphatase inhibitors, The DNA maximum absorption peak was measured at 260 nm. C Cumulative release of GEM when PTK7-GEMs were incubated with crude cell lysate, phosphatase inhibitor-treated cell lysate, buffer at pH 5.5, 10% FBS and 10 ⋅ 10 − 3 м GSH respectively

Article Snippet: Thereafter, the membranes were reacted with rabbit polyclonal antibody recognizing PTK7 (Abcam; 1:500) or ACTB (quality control, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:800) overnight at 4 °C.

Techniques: Incubation

Journal: Biomaterials Research

Article Title: Targeting treatment of bladder cancer using PTK7 aptamer-gemcitabine conjugate

doi: 10.1186/s40824-022-00328-9

Figure Lengend Snippet: PTK7-GEMs internalization and trafficking in cells. Images generated using confocal microscopy revealing the co-localization of PTK7-GEMs-cy5 (red) with markers of endocytosis dextran ( A ), choleratoxin ( B ), transferrin ( C ) labeled with Alexa Fluor 488 respectively. D Pearson’s correlation coefficient analysis of PTK7-GEMs-cy5 and endocytosis marker co-localization in 5637 cells using Image J. Confocal microscopy images revealing the co-localization of PTK7-GEMs-cy5 (red) with EIPA ( E ) (inhibitor of macropinocytosis), Filipin ( F ) (inhibitor of the caveolae pathway), and Chlorpromazine ( G ) (inhibitor of the clathrin pathway) (green). DAPI was used to counterstain the nuclei (blue). Scale bar, 10 μm. The means ± standard deviation ( n = 3 per group) is indicated using error bars. Each replicate was from a single biological experiment and 10 independent fields of view were chosen from quantification. * P < 0.05; NS = not significant

Article Snippet: Thereafter, the membranes were reacted with rabbit polyclonal antibody recognizing PTK7 (Abcam; 1:500) or ACTB (quality control, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:800) overnight at 4 °C.

Techniques: Generated, Confocal Microscopy, Labeling, Marker, Standard Deviation

Journal: Biomaterials Research

Article Title: Targeting treatment of bladder cancer using PTK7 aptamer-gemcitabine conjugate

doi: 10.1186/s40824-022-00328-9

Figure Lengend Snippet: Optical imaging and Anti-tumor activity of PTK7-GEMs in xenotransplanted subcutaneous tumors. A Time-lapse fluorescence imaging at 0.5, 1, 2, 3, and 4 h after tail vein injection of Cy5-labeled PTK7-GEMs (right) or Lib-GEMs (left). The tumor site is indicated using a red circle. B Biodistribution of PTK7-GEMs-cy5 or Lib-GEMs-cy5 in the major viscera and tumors at 4 h after injection. C Growth curve of the tumors during therapy. In vivo evaluation of antitumor effects in the 5637 xenografted bladder cancer mouse model ( n = 6 mice per group, n = 6 groups). D The tumor weights from the different groups are shown as a scattergram. E Images of tumor tissues from mice sacrificed at 32 days post inoculation. F Body weight changes during therapy. Data represents the mean ± SEM, n = 6; NS = not significant. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: Thereafter, the membranes were reacted with rabbit polyclonal antibody recognizing PTK7 (Abcam; 1:500) or ACTB (quality control, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:800) overnight at 4 °C.

Techniques: Optical Imaging, Activity Assay, Fluorescence, Imaging, Injection, Labeling, In Vivo

Journal: Biomaterials Research

Article Title: Targeting treatment of bladder cancer using PTK7 aptamer-gemcitabine conjugate

doi: 10.1186/s40824-022-00328-9

Figure Lengend Snippet: The antitumor activity of PTK7-GEMs in a mouse model of lung metastasis bladder cancer induced by tail vein injection. A Photographs of lungs excised from mice bearing bladder cancer cell tumors treated as indicated. B Photographs of lung tissue sections from mice bearing bladder cancer cell tumors under different treatments stained with H&E. C Comparison of pulmonary metastatic nodules of bladder cancer cells tumor-bearing mice treated as indicated. D Changes in the body weights of bladder cancer cells bearing mice under various treatments. All data are presented as the mean ± SD ( n = 5). ** P < 0.01, *** P < 0.001

Article Snippet: Thereafter, the membranes were reacted with rabbit polyclonal antibody recognizing PTK7 (Abcam; 1:500) or ACTB (quality control, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:800) overnight at 4 °C.

Techniques: Activity Assay, Injection, Staining, Comparison

Journal: Biomaterials Research

Article Title: Targeting treatment of bladder cancer using PTK7 aptamer-gemcitabine conjugate

doi: 10.1186/s40824-022-00328-9

Figure Lengend Snippet: The antitumor activity of PTK7-GEMs in a model of in-situ bladder cancer model. A Photographs of H&E-stained tissue sections (muscle invasive bladder cancer at stage pT2 or higher and excised bladders after treatment with PBS. B Photographs of H&E-stained tissue sections (muscle invasive bladder cancer at stage pT2 or higher and excised bladders after treatment with PTK7. ( C ) Photographs of H&E-stained tissue sections (muscle invasive bladder cancer at stage pT2 or higher and excised bladders after treatment with LIB. D Photographs of H&E-stained tissue sections (noninvasive papillary carcinoma: stage pT1) and excised bladders after treatment with GEM. E Photographs of H&E-stained tissue sections (noninvasive papillary carcinoma: stage pT1) and excised bladders after treatment with LIB-GEMs. F Photographs of H&E-stained tissue sections (noninvasive papillary carcinoma: at stage pTa or lower) and excided bladders after treatment with PTK7-GEMs.

Article Snippet: Thereafter, the membranes were reacted with rabbit polyclonal antibody recognizing PTK7 (Abcam; 1:500) or ACTB (quality control, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:800) overnight at 4 °C.

Techniques: Activity Assay, In Situ, Staining

Journal: Molecular cell

Article Title: Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene

doi: 10.1016/j.molcel.2017.11.004

Figure Lengend Snippet: (A) AMIGO2 expression levels detected by qRT-PCR in NHM and a panel of metastatic melanoma cell lines harboring distinct melanoma mutations. Data are normalized to GAPDH.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies BRD2 Bethyl Laboratories A302-583A BRD4 Bethyl Laboratories A301-985A H3K4me1 Abcam ab8895 H3K4me3 Abcam ab1012 H3K27ac Abcam ab177178 H3K27me3 EMD 07-449 MED1 Bethyl Laboratories A300-793A macroH2A2 Bernstein Laboratory, ISMMS in house macroH2A1 Abcam ab37264 CTCF EMD 07-729 FOSL2 Santa Cruz SC-604 TEAD4 Santa Cruz SC-101184 HSP90 Santa Cruz SC-13119 GAPDH Santa Cruz SC-32233 GFP Roche 11814460001 LAMIN B1 Santa Cruz SC-6217 ALCAM Abcam ab109215 L1CAM Abcam ab20148 NRP2 Santa Cruz SC-13117 NRP1 Abcam ab81321 TUBULIN Sigma T9026 FOXM1 Cell Signaling Technology 5436 Casp3 Cell Signaling Technology 9662 PARP Cell Signaling Technology 9542 AMIGO2 (for IHC) Abnova H00347902-B01P Lot# F8271 RRID:AB_1672842 AMIGO2 (for IF and WB) R&D systems AF2080 PTK7 (N-terminal) (for WB) R&D systems AF4499 PTK7 (C-terminal) (for IF and WB) Cell Signaling 11926 Secondary rabbit anti goat (for AMIGO2 detection on WB) Sigma A5420 HRP-conjugated ACTIN Sigma A3854 Biological Samples Melanoma and normal skin tissue microarray Biomax ME208 Nevus tissue microarray IMCG, NYU MT032808 Patient derived short term cultures Osman Laboratory, NYU de Miera et al., 2012 NHM (this paper) Bernstein Laboratory, ISMMS NHM1-5 Chemicals, Peptides, and Recombinant Proteins GFP Trap-A beads Chromotek gta-10 BET inhibitor JQ1 Bradner Laboratory; Agreement # {"type":"entrez-nucleotide","attrs":{"text":"A09608","term_id":"490581","term_text":"A09608"}} A09608 N/A BET inhibitor I-BET762 Apex Bio 501014747 IGEPAL Sigma I-8896 Micrococcal Nuclease Affymetrix 70196Y Dynabeads Protein A Immunoprécipitation kit Invitrogen 10006D Magna ChIP Protein A+G Magnetic beads Millipore 16-663 DSG Pierce (ThermoFisher) 20593 Critical Commercial Assays Click- Click-IT EdU Pacific Blue Flow Cytometry Assay Kit ThermoFisher {"type":"entrez-nucleotide","attrs":{"text":"C10636","term_id":"1535707","term_text":"C10636"}} C10636 Caspase Activity assay MBL Int.

Techniques: Expressing, Quantitative RT-PCR

Journal: Molecular cell

Article Title: Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene

doi: 10.1016/j.molcel.2017.11.004

Figure Lengend Snippet: (A) Relative growth curves of 501MEL and SKmel147 cells stably transduced with non-targeting scrambled control shRNA (shSCR) and two AMIGO2 shRNAs (shA2 #1 and #2). Values are normalized to seeding control (n = 3).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies BRD2 Bethyl Laboratories A302-583A BRD4 Bethyl Laboratories A301-985A H3K4me1 Abcam ab8895 H3K4me3 Abcam ab1012 H3K27ac Abcam ab177178 H3K27me3 EMD 07-449 MED1 Bethyl Laboratories A300-793A macroH2A2 Bernstein Laboratory, ISMMS in house macroH2A1 Abcam ab37264 CTCF EMD 07-729 FOSL2 Santa Cruz SC-604 TEAD4 Santa Cruz SC-101184 HSP90 Santa Cruz SC-13119 GAPDH Santa Cruz SC-32233 GFP Roche 11814460001 LAMIN B1 Santa Cruz SC-6217 ALCAM Abcam ab109215 L1CAM Abcam ab20148 NRP2 Santa Cruz SC-13117 NRP1 Abcam ab81321 TUBULIN Sigma T9026 FOXM1 Cell Signaling Technology 5436 Casp3 Cell Signaling Technology 9662 PARP Cell Signaling Technology 9542 AMIGO2 (for IHC) Abnova H00347902-B01P Lot# F8271 RRID:AB_1672842 AMIGO2 (for IF and WB) R&D systems AF2080 PTK7 (N-terminal) (for WB) R&D systems AF4499 PTK7 (C-terminal) (for IF and WB) Cell Signaling 11926 Secondary rabbit anti goat (for AMIGO2 detection on WB) Sigma A5420 HRP-conjugated ACTIN Sigma A3854 Biological Samples Melanoma and normal skin tissue microarray Biomax ME208 Nevus tissue microarray IMCG, NYU MT032808 Patient derived short term cultures Osman Laboratory, NYU de Miera et al., 2012 NHM (this paper) Bernstein Laboratory, ISMMS NHM1-5 Chemicals, Peptides, and Recombinant Proteins GFP Trap-A beads Chromotek gta-10 BET inhibitor JQ1 Bradner Laboratory; Agreement # {"type":"entrez-nucleotide","attrs":{"text":"A09608","term_id":"490581","term_text":"A09608"}} A09608 N/A BET inhibitor I-BET762 Apex Bio 501014747 IGEPAL Sigma I-8896 Micrococcal Nuclease Affymetrix 70196Y Dynabeads Protein A Immunoprécipitation kit Invitrogen 10006D Magna ChIP Protein A+G Magnetic beads Millipore 16-663 DSG Pierce (ThermoFisher) 20593 Critical Commercial Assays Click- Click-IT EdU Pacific Blue Flow Cytometry Assay Kit ThermoFisher {"type":"entrez-nucleotide","attrs":{"text":"C10636","term_id":"1535707","term_text":"C10636"}} C10636 Caspase Activity assay MBL Int.

Techniques: Stable Transfection, Transduction, shRNA

Journal: Molecular cell

Article Title: Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene

doi: 10.1016/j.molcel.2017.11.004

Figure Lengend Snippet: (A) Immunofluorescence of SKmel147 cells stably expressing AMIGO2-GFP (green), stained with AMIGO2 antibody (red) and Hoechst 33342 (blue). Scale bar, 20 μm.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies BRD2 Bethyl Laboratories A302-583A BRD4 Bethyl Laboratories A301-985A H3K4me1 Abcam ab8895 H3K4me3 Abcam ab1012 H3K27ac Abcam ab177178 H3K27me3 EMD 07-449 MED1 Bethyl Laboratories A300-793A macroH2A2 Bernstein Laboratory, ISMMS in house macroH2A1 Abcam ab37264 CTCF EMD 07-729 FOSL2 Santa Cruz SC-604 TEAD4 Santa Cruz SC-101184 HSP90 Santa Cruz SC-13119 GAPDH Santa Cruz SC-32233 GFP Roche 11814460001 LAMIN B1 Santa Cruz SC-6217 ALCAM Abcam ab109215 L1CAM Abcam ab20148 NRP2 Santa Cruz SC-13117 NRP1 Abcam ab81321 TUBULIN Sigma T9026 FOXM1 Cell Signaling Technology 5436 Casp3 Cell Signaling Technology 9662 PARP Cell Signaling Technology 9542 AMIGO2 (for IHC) Abnova H00347902-B01P Lot# F8271 RRID:AB_1672842 AMIGO2 (for IF and WB) R&D systems AF2080 PTK7 (N-terminal) (for WB) R&D systems AF4499 PTK7 (C-terminal) (for IF and WB) Cell Signaling 11926 Secondary rabbit anti goat (for AMIGO2 detection on WB) Sigma A5420 HRP-conjugated ACTIN Sigma A3854 Biological Samples Melanoma and normal skin tissue microarray Biomax ME208 Nevus tissue microarray IMCG, NYU MT032808 Patient derived short term cultures Osman Laboratory, NYU de Miera et al., 2012 NHM (this paper) Bernstein Laboratory, ISMMS NHM1-5 Chemicals, Peptides, and Recombinant Proteins GFP Trap-A beads Chromotek gta-10 BET inhibitor JQ1 Bradner Laboratory; Agreement # {"type":"entrez-nucleotide","attrs":{"text":"A09608","term_id":"490581","term_text":"A09608"}} A09608 N/A BET inhibitor I-BET762 Apex Bio 501014747 IGEPAL Sigma I-8896 Micrococcal Nuclease Affymetrix 70196Y Dynabeads Protein A Immunoprécipitation kit Invitrogen 10006D Magna ChIP Protein A+G Magnetic beads Millipore 16-663 DSG Pierce (ThermoFisher) 20593 Critical Commercial Assays Click- Click-IT EdU Pacific Blue Flow Cytometry Assay Kit ThermoFisher {"type":"entrez-nucleotide","attrs":{"text":"C10636","term_id":"1535707","term_text":"C10636"}} C10636 Caspase Activity assay MBL Int.

Techniques: Immunofluorescence, Stable Transfection, Expressing, Staining

Journal: Molecular cell

Article Title: Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene

doi: 10.1016/j.molcel.2017.11.004

Figure Lengend Snippet: (A) Capture of the UCSC genome browser (GRCh37/hg19) showing 800 kb region on human chromosome 12 (y axis = reads per kilobase per million reads). Ensembl genes (blue) and AMIGO2 SEs (red) are shown at the top. Putative TAD/LAD domain is shown at the bottom.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies BRD2 Bethyl Laboratories A302-583A BRD4 Bethyl Laboratories A301-985A H3K4me1 Abcam ab8895 H3K4me3 Abcam ab1012 H3K27ac Abcam ab177178 H3K27me3 EMD 07-449 MED1 Bethyl Laboratories A300-793A macroH2A2 Bernstein Laboratory, ISMMS in house macroH2A1 Abcam ab37264 CTCF EMD 07-729 FOSL2 Santa Cruz SC-604 TEAD4 Santa Cruz SC-101184 HSP90 Santa Cruz SC-13119 GAPDH Santa Cruz SC-32233 GFP Roche 11814460001 LAMIN B1 Santa Cruz SC-6217 ALCAM Abcam ab109215 L1CAM Abcam ab20148 NRP2 Santa Cruz SC-13117 NRP1 Abcam ab81321 TUBULIN Sigma T9026 FOXM1 Cell Signaling Technology 5436 Casp3 Cell Signaling Technology 9662 PARP Cell Signaling Technology 9542 AMIGO2 (for IHC) Abnova H00347902-B01P Lot# F8271 RRID:AB_1672842 AMIGO2 (for IF and WB) R&D systems AF2080 PTK7 (N-terminal) (for WB) R&D systems AF4499 PTK7 (C-terminal) (for IF and WB) Cell Signaling 11926 Secondary rabbit anti goat (for AMIGO2 detection on WB) Sigma A5420 HRP-conjugated ACTIN Sigma A3854 Biological Samples Melanoma and normal skin tissue microarray Biomax ME208 Nevus tissue microarray IMCG, NYU MT032808 Patient derived short term cultures Osman Laboratory, NYU de Miera et al., 2012 NHM (this paper) Bernstein Laboratory, ISMMS NHM1-5 Chemicals, Peptides, and Recombinant Proteins GFP Trap-A beads Chromotek gta-10 BET inhibitor JQ1 Bradner Laboratory; Agreement # {"type":"entrez-nucleotide","attrs":{"text":"A09608","term_id":"490581","term_text":"A09608"}} A09608 N/A BET inhibitor I-BET762 Apex Bio 501014747 IGEPAL Sigma I-8896 Micrococcal Nuclease Affymetrix 70196Y Dynabeads Protein A Immunoprécipitation kit Invitrogen 10006D Magna ChIP Protein A+G Magnetic beads Millipore 16-663 DSG Pierce (ThermoFisher) 20593 Critical Commercial Assays Click- Click-IT EdU Pacific Blue Flow Cytometry Assay Kit ThermoFisher {"type":"entrez-nucleotide","attrs":{"text":"C10636","term_id":"1535707","term_text":"C10636"}} C10636 Caspase Activity assay MBL Int.

Techniques:

Journal: Molecular cell

Article Title: Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene

doi: 10.1016/j.molcel.2017.11.004

Figure Lengend Snippet: (A) Schematic of AMIGO2 SE showing enhancers E1–E5 (B), deleted sequences for CRISPR-Cas9 editing (C), and DNA segments used for E3 lucif-erase assay (D).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies BRD2 Bethyl Laboratories A302-583A BRD4 Bethyl Laboratories A301-985A H3K4me1 Abcam ab8895 H3K4me3 Abcam ab1012 H3K27ac Abcam ab177178 H3K27me3 EMD 07-449 MED1 Bethyl Laboratories A300-793A macroH2A2 Bernstein Laboratory, ISMMS in house macroH2A1 Abcam ab37264 CTCF EMD 07-729 FOSL2 Santa Cruz SC-604 TEAD4 Santa Cruz SC-101184 HSP90 Santa Cruz SC-13119 GAPDH Santa Cruz SC-32233 GFP Roche 11814460001 LAMIN B1 Santa Cruz SC-6217 ALCAM Abcam ab109215 L1CAM Abcam ab20148 NRP2 Santa Cruz SC-13117 NRP1 Abcam ab81321 TUBULIN Sigma T9026 FOXM1 Cell Signaling Technology 5436 Casp3 Cell Signaling Technology 9662 PARP Cell Signaling Technology 9542 AMIGO2 (for IHC) Abnova H00347902-B01P Lot# F8271 RRID:AB_1672842 AMIGO2 (for IF and WB) R&D systems AF2080 PTK7 (N-terminal) (for WB) R&D systems AF4499 PTK7 (C-terminal) (for IF and WB) Cell Signaling 11926 Secondary rabbit anti goat (for AMIGO2 detection on WB) Sigma A5420 HRP-conjugated ACTIN Sigma A3854 Biological Samples Melanoma and normal skin tissue microarray Biomax ME208 Nevus tissue microarray IMCG, NYU MT032808 Patient derived short term cultures Osman Laboratory, NYU de Miera et al., 2012 NHM (this paper) Bernstein Laboratory, ISMMS NHM1-5 Chemicals, Peptides, and Recombinant Proteins GFP Trap-A beads Chromotek gta-10 BET inhibitor JQ1 Bradner Laboratory; Agreement # {"type":"entrez-nucleotide","attrs":{"text":"A09608","term_id":"490581","term_text":"A09608"}} A09608 N/A BET inhibitor I-BET762 Apex Bio 501014747 IGEPAL Sigma I-8896 Micrococcal Nuclease Affymetrix 70196Y Dynabeads Protein A Immunoprécipitation kit Invitrogen 10006D Magna ChIP Protein A+G Magnetic beads Millipore 16-663 DSG Pierce (ThermoFisher) 20593 Critical Commercial Assays Click- Click-IT EdU Pacific Blue Flow Cytometry Assay Kit ThermoFisher {"type":"entrez-nucleotide","attrs":{"text":"C10636","term_id":"1535707","term_text":"C10636"}} C10636 Caspase Activity assay MBL Int.

Techniques: CRISPR

Journal: Molecular cell

Article Title: Harnessing BET Inhibitor Sensitivity Reveals AMIGO2 as a Melanoma Survival Gene

doi: 10.1016/j.molcel.2017.11.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies BRD2 Bethyl Laboratories A302-583A BRD4 Bethyl Laboratories A301-985A H3K4me1 Abcam ab8895 H3K4me3 Abcam ab1012 H3K27ac Abcam ab177178 H3K27me3 EMD 07-449 MED1 Bethyl Laboratories A300-793A macroH2A2 Bernstein Laboratory, ISMMS in house macroH2A1 Abcam ab37264 CTCF EMD 07-729 FOSL2 Santa Cruz SC-604 TEAD4 Santa Cruz SC-101184 HSP90 Santa Cruz SC-13119 GAPDH Santa Cruz SC-32233 GFP Roche 11814460001 LAMIN B1 Santa Cruz SC-6217 ALCAM Abcam ab109215 L1CAM Abcam ab20148 NRP2 Santa Cruz SC-13117 NRP1 Abcam ab81321 TUBULIN Sigma T9026 FOXM1 Cell Signaling Technology 5436 Casp3 Cell Signaling Technology 9662 PARP Cell Signaling Technology 9542 AMIGO2 (for IHC) Abnova H00347902-B01P Lot# F8271 RRID:AB_1672842 AMIGO2 (for IF and WB) R&D systems AF2080 PTK7 (N-terminal) (for WB) R&D systems AF4499 PTK7 (C-terminal) (for IF and WB) Cell Signaling 11926 Secondary rabbit anti goat (for AMIGO2 detection on WB) Sigma A5420 HRP-conjugated ACTIN Sigma A3854 Biological Samples Melanoma and normal skin tissue microarray Biomax ME208 Nevus tissue microarray IMCG, NYU MT032808 Patient derived short term cultures Osman Laboratory, NYU de Miera et al., 2012 NHM (this paper) Bernstein Laboratory, ISMMS NHM1-5 Chemicals, Peptides, and Recombinant Proteins GFP Trap-A beads Chromotek gta-10 BET inhibitor JQ1 Bradner Laboratory; Agreement # {"type":"entrez-nucleotide","attrs":{"text":"A09608","term_id":"490581","term_text":"A09608"}} A09608 N/A BET inhibitor I-BET762 Apex Bio 501014747 IGEPAL Sigma I-8896 Micrococcal Nuclease Affymetrix 70196Y Dynabeads Protein A Immunoprécipitation kit Invitrogen 10006D Magna ChIP Protein A+G Magnetic beads Millipore 16-663 DSG Pierce (ThermoFisher) 20593 Critical Commercial Assays Click- Click-IT EdU Pacific Blue Flow Cytometry Assay Kit ThermoFisher {"type":"entrez-nucleotide","attrs":{"text":"C10636","term_id":"1535707","term_text":"C10636"}} C10636 Caspase Activity assay MBL Int.

Techniques: Microarray, Derivative Assay, Recombinant, Magnetic Beads, Flow Cytometry, Caspase Activity Assay, Blocking Assay, TA Cloning, Sequencing, RNA Sequencing Assay, Western Blot, Mass Spectrometry, Expressing, Software