Journal: Current Pharmaceutical Biotechnology

Article Title: Evaluation of Changes in the Expression Pattern of EDIL3 in Different Grades of Endometrial Cancer

doi: 10.2174/1389201020666190408112822

Figure Lengend Snippet: Optical density of the reaction product for EDIL3 in different grades of endometrial cancer and control.

Article Snippet: Rabbit polyclonal anti-EDIL3 antibody (Novus Biological, USA) was used to perform immunohistochemical staining of the prepared slides.

Techniques: Control

Journal: Cell Death Discovery

Article Title: EDIL3 promotes epithelial–mesenchymal transition and paclitaxel resistance through its interaction with integrin α V β 3 in cancer cells

doi: 10.1038/s41420-020-00322-x

Figure Lengend Snippet: Western blot analysis of basal expression of EDIL3, and different epithelial and mesenchymal markers are shown. β-actin is shown as a loading control. Histograms show the densitometric analyses of indicated proteins. Data are presented as mean ± SEM ( n ≥ 3). a EDIL3 expression in MDA-MB-468, MDA-MB-468R, BT474, SKBR3, and MDA-MB-231 breast cancer cells. b E-cadherin, vimentin, and Slug expression in MDA-MB-468, MDA-MB-468R, and MDA-MB-231 breast cancer cells. c EDIL3, E-cadherin, vimentin, and Slug expression in LNCaP and PC3 prostate cancer cells. d , e Representative micrographs of EDIL3 immunohistochemistry in low- and high-grade breast tumors ( d ), and in low- and high-grade prostate tumors ( e ). Bars, 75 µm. Histograms show percent of breast ( d ) or prostate ( e ) tumors with high EDIL3 expression. Association between EDIL3 and tumor grade was analyzed by Fisher’s exact test. * P < 0.05 from Student’s t test; NS not significant.

Article Snippet: The antibodies and dilutions used for western blots were as follows: rabbit polyclonal anti-EDIL3 (1:1000, cat #PA5-27994, Thermo Fisher Scientific, Waltham, MA, USA), mouse monoclonal anti-β-actin (1:20000, cat #A5441, Sigma), rabbit polyclonal anti-cleaved caspase-3 (1:500, cat #9664, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal anti-Slug (1:1000, cat #166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse monoclonal anti-PARP (1:500, cat #551024, BD Biosciences), mouse monoclonal anti-E-cadherin (1:5000; cat #610181, BD Biosciences), and rabbit monoclonal anti-vimentin (1:1000, cat #92547, Abcam, Cambridge, UK).

Techniques: Western Blot, Expressing, Immunohistochemistry

Journal: Cell Death Discovery

Article Title: EDIL3 promotes epithelial–mesenchymal transition and paclitaxel resistance through its interaction with integrin α V β 3 in cancer cells

doi: 10.1038/s41420-020-00322-x

Figure Lengend Snippet: a IC50 curve for cilengitide in MDA-MB-231 breast cancer cell line. Data are presented as mean ± SEM. b Histogram shows the concentration of EDIL3 protein in the medium determined by ELISA in MDA-MB-468, MDA-MB-468R, and MDA-MB-231 breast cancer cells treated with DMSO or 1 µM cilengitide during 48 h. c Western blot analysis of EDIL3, E-cadherin, vimentin, and Slug in MDA-MB-231 cells treated with DMSO and 1 µM cilengitide during 48 h. β-actin is shown as a loading control. Histograms show the densitometric analyses of indicated proteins. Data are presented as mean ± SEM ( n ≥ 3). d Western blot analysis of EDIL3, E-cadherin, vimentin, and Slug in MDA-MB-468 cells cultured in unconditioned medium (UM) or EDIL3-enriched conditioned medium collected from MDA-MB-231 cells (CM), and treated with DMSO and 1 µM cilengitide during 48 h. β-actin is shown as a loading control. Histograms show the densitometric analyses of indicated proteins. Data are presented as mean ± SEM ( n ≥ 3). * P < 0.05 from Student’s t test; NS not significant.

Article Snippet: The antibodies and dilutions used for western blots were as follows: rabbit polyclonal anti-EDIL3 (1:1000, cat #PA5-27994, Thermo Fisher Scientific, Waltham, MA, USA), mouse monoclonal anti-β-actin (1:20000, cat #A5441, Sigma), rabbit polyclonal anti-cleaved caspase-3 (1:500, cat #9664, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal anti-Slug (1:1000, cat #166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse monoclonal anti-PARP (1:500, cat #551024, BD Biosciences), mouse monoclonal anti-E-cadherin (1:5000; cat #610181, BD Biosciences), and rabbit monoclonal anti-vimentin (1:1000, cat #92547, Abcam, Cambridge, UK).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Cell Culture

Journal: Cell Death Discovery

Article Title: EDIL3 promotes epithelial–mesenchymal transition and paclitaxel resistance through its interaction with integrin α V β 3 in cancer cells

doi: 10.1038/s41420-020-00322-x

Figure Lengend Snippet: a MDA-MB-231 and b PC3 cells were silenced for EDIL3, and treated with DMSO and 1 or 2.5 µM paclitaxel, respectively. Paclitaxel-induced apoptosis was assessed by western blot analysis of cleaved PARP and active caspase-3. Modulation of EDIL3, E-cadherin, vimentin, and Slug was studied by western blot analysis, using β-actin as a loading control. Histograms show the densitometric analyses of indicated proteins. Data are presented as mean ± SEM ( n ≥ 3). * P < 0.05 from Student’s t test; NS not significant.

Article Snippet: The antibodies and dilutions used for western blots were as follows: rabbit polyclonal anti-EDIL3 (1:1000, cat #PA5-27994, Thermo Fisher Scientific, Waltham, MA, USA), mouse monoclonal anti-β-actin (1:20000, cat #A5441, Sigma), rabbit polyclonal anti-cleaved caspase-3 (1:500, cat #9664, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal anti-Slug (1:1000, cat #166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse monoclonal anti-PARP (1:500, cat #551024, BD Biosciences), mouse monoclonal anti-E-cadherin (1:5000; cat #610181, BD Biosciences), and rabbit monoclonal anti-vimentin (1:1000, cat #92547, Abcam, Cambridge, UK).

Techniques: Western Blot

Journal: Cell Death Discovery

Article Title: EDIL3 promotes epithelial–mesenchymal transition and paclitaxel resistance through its interaction with integrin α V β 3 in cancer cells

doi: 10.1038/s41420-020-00322-x

Figure Lengend Snippet: a MDA-MB-468 breast cancer cells were cultured in unconditioned medium (UM) or EDIL3-enriched conditioned medium collected from MDA-MB-231 cells (CM), and treated with DMSO and 1 µM paclitaxel during 48 h. b LNCaP prostate cancer cells were transiently transfected with pCMV6-XL4-EDIL3 or with empty vector, and treated with DMSO or 2.5 µM paclitaxel during 48 h. Paclitaxel-induced cleavage of PARP and caspase-3, as well as paclitaxel-induced modulation of EDIL3, E-cadherin, vimentin, and Slug were analyzed by western blot using β-actin as a loading control. Histograms represent the densitometric analyses of indicated proteins. Data are presented as mean ± SEM ( n ≥ 3). * P < 0.05 from Student’s t test; NS, not significant.

Article Snippet: The antibodies and dilutions used for western blots were as follows: rabbit polyclonal anti-EDIL3 (1:1000, cat #PA5-27994, Thermo Fisher Scientific, Waltham, MA, USA), mouse monoclonal anti-β-actin (1:20000, cat #A5441, Sigma), rabbit polyclonal anti-cleaved caspase-3 (1:500, cat #9664, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal anti-Slug (1:1000, cat #166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse monoclonal anti-PARP (1:500, cat #551024, BD Biosciences), mouse monoclonal anti-E-cadherin (1:5000; cat #610181, BD Biosciences), and rabbit monoclonal anti-vimentin (1:1000, cat #92547, Abcam, Cambridge, UK).

Techniques: Cell Culture, Transfection, Plasmid Preparation, Western Blot

Journal: Cell Death Discovery

Article Title: EDIL3 promotes epithelial–mesenchymal transition and paclitaxel resistance through its interaction with integrin α V β 3 in cancer cells

doi: 10.1038/s41420-020-00322-x

Figure Lengend Snippet: a MDA-MB-231 and b PC3 cells were treated with DMSO, 1 µM cilengitide, 1 or 2.5 µM paclitaxel, and 1 µM cilengitide plus 1 or 2.5 µM paclitaxel, respectively. The induction of apoptosis was studied through the cleavage of PARP and caspase-3 by western blot, as well as the expression of indicated epithelial–mesenchymal transition markers. β-actin was used as a loading control. Histograms show the densitometric analyses of indicated proteins. Data are presented as mean ± SEM ( n ≥ 3). * P < 0.05 from Student’s t test; NS not significant.

Article Snippet: The antibodies and dilutions used for western blots were as follows: rabbit polyclonal anti-EDIL3 (1:1000, cat #PA5-27994, Thermo Fisher Scientific, Waltham, MA, USA), mouse monoclonal anti-β-actin (1:20000, cat #A5441, Sigma), rabbit polyclonal anti-cleaved caspase-3 (1:500, cat #9664, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal anti-Slug (1:1000, cat #166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse monoclonal anti-PARP (1:500, cat #551024, BD Biosciences), mouse monoclonal anti-E-cadherin (1:5000; cat #610181, BD Biosciences), and rabbit monoclonal anti-vimentin (1:1000, cat #92547, Abcam, Cambridge, UK).

Techniques: Western Blot, Expressing