Journal: Cytometry

Article Title: Quantitative study of dynamic behavior of cell monolayers during in vitro wound healing by optical flow analysis

doi: 10.1002/1097-0320(20000901)41:1<19::aid-cyto3>3.0.co;2-x

Figure Lengend Snippet: FIG. 1. Phase contrast images showing the different textures and as- pects of HeLa cells (bar 5 50 mm), L929 (bar 5 50 mm), and Rbdo (bar 5 25 mm) cells during the wound-healing process.

Article Snippet: Experiments were performed on three different cell lines: L929 murine fibroblasts (ATCC, CCL 1), human epithelioid HeLa cells (ATCC, CCL 2.2), and rat rhabdomyosarcoma (Rbdo) cells (kindly provided by Dr. M.F.

Techniques:

Journal: Cytometry

Article Title: Quantitative study of dynamic behavior of cell monolayers during in vitro wound healing by optical flow analysis

doi: 10.1002/1097-0320(20000901)41:1<19::aid-cyto3>3.0.co;2-x

Figure Lengend Snippet: FIG. 3. Evolution with time of translation velocities at the wound margins for HeLa cells. A: Just after wound scratching, vx (left and right) increased, then stabilized before decreasing after 30 h. At that time, the wound area closure was about 75%. B: Except immediately after wound scratching, the velocity vy (left and right) was small compared to vx (left and right) at the same time, indicating that HeLa cell movement occurred mostly perpendicular to the wound margin.

Article Snippet: Experiments were performed on three different cell lines: L929 murine fibroblasts (ATCC, CCL 1), human epithelioid HeLa cells (ATCC, CCL 2.2), and rat rhabdomyosarcoma (Rbdo) cells (kindly provided by Dr. M.F.

Techniques:

Journal: Cytometry

Article Title: Quantitative study of dynamic behavior of cell monolayers during in vitro wound healing by optical flow analysis

doi: 10.1002/1097-0320(20000901)41:1<19::aid-cyto3>3.0.co;2-x

Figure Lengend Snippet: FIG. 4. A: Decrease over time of the relative wound areas during the wound closure process of HeLa cells. The areas Ax(t) and As(t) were compared to Ae(t). Considering only the vx (left or right) velocity com- ponent gave a slightly underestimated area Ax(t). B: Evolution with time of the associated velocities Vfx(t), VfT(t), and Ve(t).

Article Snippet: Experiments were performed on three different cell lines: L929 murine fibroblasts (ATCC, CCL 1), human epithelioid HeLa cells (ATCC, CCL 2.2), and rat rhabdomyosarcoma (Rbdo) cells (kindly provided by Dr. M.F.

Techniques:

Journal: Cytometry

Article Title: Quantitative study of dynamic behavior of cell monolayers during in vitro wound healing by optical flow analysis

doi: 10.1002/1097-0320(20000901)41:1<19::aid-cyto3>3.0.co;2-x

Figure Lengend Snippet: FIG. 5. Evolution of the vx (left and right) velocity profile over the HeLa cell monolayer obtained using the sliding window procedure (see Mate- rials and Methods). Only cells located no further than 200 mm moved toward the center and thus actively took part in the wound-healing process.

Article Snippet: Experiments were performed on three different cell lines: L929 murine fibroblasts (ATCC, CCL 1), human epithelioid HeLa cells (ATCC, CCL 2.2), and rat rhabdomyosarcoma (Rbdo) cells (kindly provided by Dr. M.F.

Techniques:

Journal: Cytometry

Article Title: Quantitative study of dynamic behavior of cell monolayers during in vitro wound healing by optical flow analysis

doi: 10.1002/1097-0320(20000901)41:1<19::aid-cyto3>3.0.co;2-x

Figure Lengend Snippet: FIG. 6. Decrease over time of the relative wound areas Ax(t), As(t), and Ae(t) during the wound-closure process of L929. The wound closure was slower than in the case of HeLa cells (Fig. 4), even if the curves retained similar features. However, the increasing difference between As(t) and Ax(t) indicated that the vy (left or right) velocity of fibroblasts at the wound margin was raised.

Article Snippet: Experiments were performed on three different cell lines: L929 murine fibroblasts (ATCC, CCL 1), human epithelioid HeLa cells (ATCC, CCL 2.2), and rat rhabdomyosarcoma (Rbdo) cells (kindly provided by Dr. M.F.

Techniques:

Journal: Cytometry

Article Title: Quantitative study of dynamic behavior of cell monolayers during in vitro wound healing by optical flow analysis

doi: 10.1002/1097-0320(20000901)41:1<19::aid-cyto3>3.0.co;2-x

Figure Lengend Snippet: FIG. 9. Evolution with time of the relative wound area for normal or Ho 342 stained HeLa cells. After approximately 20 h, a shift between the curves appeared, resulting from a decrease of the stained cell velocity.

Article Snippet: Experiments were performed on three different cell lines: L929 murine fibroblasts (ATCC, CCL 1), human epithelioid HeLa cells (ATCC, CCL 2.2), and rat rhabdomyosarcoma (Rbdo) cells (kindly provided by Dr. M.F.

Techniques: Staining

Journal: Cytometry

Article Title: Quantitative study of dynamic behavior of cell monolayers during in vitro wound healing by optical flow analysis

doi: 10.1002/1097-0320(20000901)41:1<19::aid-cyto3>3.0.co;2-x

Figure Lengend Snippet: FIG. 10. Phase contrast images of wounded HeLa and L929 cell monolayer and representations of the translation velocity vx (left and right) over the cell monolayer during the wound-healing process. The times T1, T2, and T3 correspond to time intervals 5 h/7.5 h, 15 h/17.5 h, and 25 h/27.5 h after wound scratching. Symbols are: from 0–1.5 mm/h, from 1.5–3 mm/h, from 3–4.5 mm/h, and I .4.5 mm/h.

Article Snippet: Experiments were performed on three different cell lines: L929 murine fibroblasts (ATCC, CCL 1), human epithelioid HeLa cells (ATCC, CCL 2.2), and rat rhabdomyosarcoma (Rbdo) cells (kindly provided by Dr. M.F.

Techniques:

Journal: Scientific Reports

Article Title: Danazol mediates collateral sensitivity via STAT3/Myc related pathway in multidrug-resistant cancer cells

doi: 10.1038/s41598-019-48169-2

Figure Lengend Snippet: Selectivity Index of Danazol on several multidrug-resistant cancer cell lines.

Article Snippet: Parental cancer cell lines, including human cervical epithelioid carcinoma HeLaS3, human hepatoblastoma HepG2, and human non-small cell lung carcinoma NCI-H460, were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques:

Journal: Scientific Reports

Article Title: Danazol mediates collateral sensitivity via STAT3/Myc related pathway in multidrug-resistant cancer cells

doi: 10.1038/s41598-019-48169-2

Figure Lengend Snippet: Cytotoxicity of natural and synthetic steroid hormones on HeLaS3 (parental) and KB/VIN (MDR) cancer cell lines.

Article Snippet: Parental cancer cell lines, including human cervical epithelioid carcinoma HeLaS3, human hepatoblastoma HepG2, and human non-small cell lung carcinoma NCI-H460, were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques:

Journal: Scientific Reports

Article Title: Danazol mediates collateral sensitivity via STAT3/Myc related pathway in multidrug-resistant cancer cells

doi: 10.1038/s41598-019-48169-2

Figure Lengend Snippet: Results of ABCB1 gene expression analysis. HeLaS3 and KB/VIN were treated with danazol for 24, 48, and 72 hours. Total RNA was extracted and ABCB1 gene expression level of each sample was quantified by real-time PCR. The ABCB1 gene expression was significantly down-regulated by danazol treatment in KB/VIN cell line. Statistical differences were evaluated by ANOVA followed post hoc analysis (Tukey’s test). * Indicates p value < 0.05 compared with control group. Data presented as mean ± SE of at least three experiments, each in triplicate.

Article Snippet: Parental cancer cell lines, including human cervical epithelioid carcinoma HeLaS3, human hepatoblastoma HepG2, and human non-small cell lung carcinoma NCI-H460, were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Control

Journal: Scientific Reports

Article Title: Danazol mediates collateral sensitivity via STAT3/Myc related pathway in multidrug-resistant cancer cells

doi: 10.1038/s41598-019-48169-2

Figure Lengend Snippet: Results of cell cycle analysis in HeLaS3 and MDR KB/VIN cells. HeLaS3 and KB/VIN were treated with culture medium ( a ), DMSO ( b ), paclitaxel ( c ; positive control), danazol ( d ), β-estradiol ( e ), and deoxycorticosterone ( f ) for 24, 48, and 72 hours. DNA contents and cell cycle distribution of each sample were determined by PI solution (X-axis PE). Danazol, β-estradiol, and deoxycorticosterone arrested KB/VIN cell at G2/M phase and caused apoptosis (increased sub G1) in a time-dependent manner.

Article Snippet: Parental cancer cell lines, including human cervical epithelioid carcinoma HeLaS3, human hepatoblastoma HepG2, and human non-small cell lung carcinoma NCI-H460, were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Cell Cycle Assay, Positive Control

Journal: Scientific Reports

Article Title: Danazol mediates collateral sensitivity via STAT3/Myc related pathway in multidrug-resistant cancer cells

doi: 10.1038/s41598-019-48169-2

Figure Lengend Snippet: Results of caspase activity and ROS levels detection assay in HeLaS3 and MDR KB/VIN cells. HeLaS3 and KB/VIN cells were treated with or without danazol for 72 hours. The activities of caspase-8 ( a ) and caspase-9 ( b ) were evaluated by Cell Meter™ Caspase Activity Apoptosis Assay Kits. Danazol activated caspase-8 in KB/VIN cells while the level remained the same in HeLaS3 cells compared with respective control group. ( c ) The treatment of danazol increased the ROS level in both HeLaS3 and KB/VIN cell lines. Menadione, a ROS inducer, was used as a positive control. Statistical differences were evaluated by the ANOVA followed post hoc analysis (Tukey’s test). * indicates p value < 0.05 compared with control group. Data presented as mean ± SE of at least two experiments, each in duplicate.

Article Snippet: Parental cancer cell lines, including human cervical epithelioid carcinoma HeLaS3, human hepatoblastoma HepG2, and human non-small cell lung carcinoma NCI-H460, were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Activity Assay, Detection Assay, Apoptosis Assay, Control, Positive Control

Journal: Scientific Reports

Article Title: Danazol mediates collateral sensitivity via STAT3/Myc related pathway in multidrug-resistant cancer cells

doi: 10.1038/s41598-019-48169-2

Figure Lengend Snippet: MYC signaling mediates danazol-induced G2/M arrest in MDR KB/VIN cells. HeLaS3 and KB/VIN were treated with or without danazol for 72 hours. The whole-treated cells were extracted and analyzed by ELISA using antibodies specific to human ( a ) STAT3-pY705, ( b ) cMYC, ( c ) CDC25, ( d ) CDK1, and ( e ) p21. The results showed danazol reduced the concentration of MYC (c-Myc) and its upstream protein STAT3-pY705 in MDR KB/VIN cells. In addition, decreases in downstream signaling proteins, CDC25 and CDK1, were shown. Each experiment was performed in duplicate. Student’s t-test, *p < 0.05 compared with cells treated with medium only control.

Article Snippet: Parental cancer cell lines, including human cervical epithelioid carcinoma HeLaS3, human hepatoblastoma HepG2, and human non-small cell lung carcinoma NCI-H460, were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Control

Journal: The Journal of biological chemistry

Article Title: Identification of a membrane-associated inhibitor(s) of epidermal growth factor-induced signal transducer and activator of transcription activation.

doi: 10.1074/jbc.273.29.18198

Figure Lengend Snippet: FIG. 1. In vivo activation of STAT proteins by EGF is observed in A431 cells but not in many other cell lines. A431, HeLa S3, and PC12 cells were deprived of serum for 17 h and then stimulated with 1 ng/ml IFN-g, 100 ng/ml EGF, or 50 ng/ml NGF for 15 min. One ml of the whole cell extract was subjected to EMSA using 32P-end-labeled M67- SIE as a probe. Three types of the STAT protein-SIE complexes (SIF-A, B, and C) were observed. SIF-A, STAT3 homodimer; SIF-B, STAT1/ STAT3 heterodimer; SIF-C, STAT1 homodimer (for details, see Refs. 30–32).

Article Snippet: Tissue Culture Cells—A431 human epidermoid carcinoma cells (ATCC, CRL-1555) and HeLa S3 human epitheloid carcinoma cells (ATCC, CCL-2.2) were cultured in Dulbecco’s modified Eagle’s medium (Life Technologies, Inc.) containing 5% bovine calf serum; PC12 rat adrenal pheochromocytoma cells (ATCC, CRL-1721) in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum, 10% horse serum, and 1 mM L-glutamine; HepG2 human hepatoma cells (ATCC, HB-8065) in Dulbecco’s modified Eagle’s medium containing 10% fetal * This project was supported in part by National Institutes of Health Grant RO1AI34522.

Techniques: In Vivo, Activation Assay, Labeling

Journal: The Journal of biological chemistry

Article Title: Identification of a membrane-associated inhibitor(s) of epidermal growth factor-induced signal transducer and activator of transcription activation.

doi: 10.1074/jbc.273.29.18198

Figure Lengend Snippet: FIG. 4. A, titration of the HeLa S3 membrane to inhibit the in vitro activation of STAT proteins in the A431 cytosol by the A431 membrane. The STAT proteins in the A431 cytosol (cyto.) fraction (2 mg protein) were activated in vitro by the A431 membrane (memb.) fraction (0.3 mg) in the presence of various protein concentrations of the HeLa S3 mem- brane fraction. The formation of the STAT1 homodimer-SIE complex (SIF-C) was observed on EMSA using 32P-end-labeled M67-SIE as a probe. B, titration of the HeLa S3 membrane to inhibit the in vitro activation of STAT proteins in the HeLa S3 cytosol by the A431 mem- brane. The STAT proteins in the HeLa S3 cytosol fraction (2 mg of protein) were activated in vitro by the A431 membrane fraction (0.3 mg) in the presence of various protein concentrations of the HeLa S3 mem- brane fraction. The formation of the STAT1 homodimer-SIE complex (SIF-C) was observed on EMSA using 32P-end-labeled M67-SIE as a probe.

Article Snippet: Tissue Culture Cells—A431 human epidermoid carcinoma cells (ATCC, CRL-1555) and HeLa S3 human epitheloid carcinoma cells (ATCC, CCL-2.2) were cultured in Dulbecco’s modified Eagle’s medium (Life Technologies, Inc.) containing 5% bovine calf serum; PC12 rat adrenal pheochromocytoma cells (ATCC, CRL-1721) in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum, 10% horse serum, and 1 mM L-glutamine; HepG2 human hepatoma cells (ATCC, HB-8065) in Dulbecco’s modified Eagle’s medium containing 10% fetal * This project was supported in part by National Institutes of Health Grant RO1AI34522.

Techniques: Titration, Membrane, In Vitro, Activation Assay, Labeling

Journal: The Journal of biological chemistry

Article Title: Identification of a membrane-associated inhibitor(s) of epidermal growth factor-induced signal transducer and activator of transcription activation.

doi: 10.1074/jbc.273.29.18198

Figure Lengend Snippet: FIG. 5. The inhibitor in HeLa S3 or PC12 membrane inhibits the in vitro activation of the STAT1 but not the binding of the activated STAT1 with SIE. Lanes 1–3 and 6, the STAT proteins in the A431 cytosol (cyto.) fraction (5.5 mg of protein) were activated in vitro by the A431 membrane (memb.) fraction (0.44 mg) in the absence or the presence of the HeLa S3 or the PC12 membrane fraction (6.2 mg). Three ml of the in vitro activated sample was subjected to EMSA using 32P- end-labeled M67-SIE as a probe. Lanes 4 and 5 and 7 and 8, two ml of buffer A, the HeLa S3, or the PC12 fraction (6.2 mg) was mixed with the in vitro activated sample (10 ml), and 3 ml of the mixture was subjected to EMSA. The formation of the STAT1 homodimer-SIE complex (SIF-C) was observed.

Article Snippet: Tissue Culture Cells—A431 human epidermoid carcinoma cells (ATCC, CRL-1555) and HeLa S3 human epitheloid carcinoma cells (ATCC, CCL-2.2) were cultured in Dulbecco’s modified Eagle’s medium (Life Technologies, Inc.) containing 5% bovine calf serum; PC12 rat adrenal pheochromocytoma cells (ATCC, CRL-1721) in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum, 10% horse serum, and 1 mM L-glutamine; HepG2 human hepatoma cells (ATCC, HB-8065) in Dulbecco’s modified Eagle’s medium containing 10% fetal * This project was supported in part by National Institutes of Health Grant RO1AI34522.

Techniques: Membrane, In Vitro, Activation Assay, Binding Assay, Labeling

Journal: The Journal of biological chemistry

Article Title: Identification of a membrane-associated inhibitor(s) of epidermal growth factor-induced signal transducer and activator of transcription activation.

doi: 10.1074/jbc.273.29.18198

Figure Lengend Snippet: FIG. 6. Partial purification of the inhibitor. A, solubilization of the STAT inhibitor from the membrane fraction. The STAT proteins in the A431 cytosol fraction (2 mg of protein) were activated in vitro by the A431 membrane fraction (0.3 mg) in the presence of HeLa S3 fractions (40 mg). The 10,000g pellet refers to the membrane fraction. The forma- tion of the STAT1 homodimer-SIE complex (SIF-C) was observed on EMSA using 32P-end-labeled M67-SIE as a probe. sup, supernatant. B,

Article Snippet: Tissue Culture Cells—A431 human epidermoid carcinoma cells (ATCC, CRL-1555) and HeLa S3 human epitheloid carcinoma cells (ATCC, CCL-2.2) were cultured in Dulbecco’s modified Eagle’s medium (Life Technologies, Inc.) containing 5% bovine calf serum; PC12 rat adrenal pheochromocytoma cells (ATCC, CRL-1721) in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum, 10% horse serum, and 1 mM L-glutamine; HepG2 human hepatoma cells (ATCC, HB-8065) in Dulbecco’s modified Eagle’s medium containing 10% fetal * This project was supported in part by National Institutes of Health Grant RO1AI34522.

Techniques: Purification, Membrane, In Vitro, Labeling

Journal: The Journal of biological chemistry

Article Title: Identification of a membrane-associated inhibitor(s) of epidermal growth factor-induced signal transducer and activator of transcription activation.

doi: 10.1074/jbc.273.29.18198

Figure Lengend Snippet: FIG. 7. STAT1 inhibitor activity in various cell lines. The STAT proteins in the A431 cytosol fraction (5.5 mg of protein) were activated in vitro by the A431 membrane (memb.) fraction (0.44 mg) in the pres- ence of the HeLa S3, PC12, HepG2, THP-1, T-47D, WiDr, HT-29, SW1116, or A431 membrane fraction (6.2 mg). Three ml of the in vitro activated sample were subjected to EMSA using 32P-end-labeled M67- SIE as a probe. The formation of the STAT1 homodimer-SIE complex (SIF-C) was observed.

Article Snippet: Tissue Culture Cells—A431 human epidermoid carcinoma cells (ATCC, CRL-1555) and HeLa S3 human epitheloid carcinoma cells (ATCC, CCL-2.2) were cultured in Dulbecco’s modified Eagle’s medium (Life Technologies, Inc.) containing 5% bovine calf serum; PC12 rat adrenal pheochromocytoma cells (ATCC, CRL-1721) in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum, 10% horse serum, and 1 mM L-glutamine; HepG2 human hepatoma cells (ATCC, HB-8065) in Dulbecco’s modified Eagle’s medium containing 10% fetal * This project was supported in part by National Institutes of Health Grant RO1AI34522.

Techniques: Activity Assay, In Vitro, Membrane, Labeling