shRNA:Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation
Article Snippet: In order to maintain the stable expression of PMCA4b transgenes and of the shRNA construct, 100 ng/ml puromycin dihydrochloride (sc-108071, Santa Cruz Biotechnology) was added to the culture media. .. PMCA4-specific shRNA MCF-7 cells were generated by using the PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA), as described previously ( Naffa et al ., 2021 ). ..
Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation.
Article Snippet: .. Generation of stable cell lines PMCA4-specific shRNA expressing MCF-7 cells were generated by transfecting with PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA) that contains puromycin resistance Fig. 8 | PMCA4b promotes central lumen formation in 3D cultures of MCF7 cells. a Phalloidin staining of mammospheres formed by parental, PMCA4specific shRNA (sh-PMCA4), GFP-PMCA4b and GFP-PMCA4bLA-expressing MCF-7 cell lines cultured inMatrigel for 10 days. b Schematic figures demonstrating structures of mammospheres based on results on (a). c Statistical comparisons of the ratios of mammospheres with central lumen in 3D cultures between parental, PMCA4-specific shRNA (sh-PMCA4), GFP-PMCA4b- and GFP-PMCA4bLAexpressing MCF-7 cell lines cultured in Matrigel. ..
Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation
Article Snippet: .. PMCA4-specific shRNA expressing MCF-7 cells were generated by transfecting with PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA) that contains puromycin resistance gene using FuGENE HD transfection reagent (Promega, Madison, WI, USA) according to the manufacturer’s instructions. ..
Generated:Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation
Article Snippet: In order to maintain the stable expression of PMCA4b transgenes and of the shRNA construct, 100 ng/ml puromycin dihydrochloride (sc-108071, Santa Cruz Biotechnology) was added to the culture media. .. PMCA4-specific shRNA MCF-7 cells were generated by using the PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA), as described previously ( Naffa et al ., 2021 ). ..
Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation.
Article Snippet: .. Generation of stable cell lines PMCA4-specific shRNA expressing MCF-7 cells were generated by transfecting with PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA) that contains puromycin resistance Fig. 8 | PMCA4b promotes central lumen formation in 3D cultures of MCF7 cells. a Phalloidin staining of mammospheres formed by parental, PMCA4specific shRNA (sh-PMCA4), GFP-PMCA4b and GFP-PMCA4bLA-expressing MCF-7 cell lines cultured inMatrigel for 10 days. b Schematic figures demonstrating structures of mammospheres based on results on (a). c Statistical comparisons of the ratios of mammospheres with central lumen in 3D cultures between parental, PMCA4-specific shRNA (sh-PMCA4), GFP-PMCA4b- and GFP-PMCA4bLAexpressing MCF-7 cell lines cultured in Matrigel. ..
Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation
Article Snippet: .. PMCA4-specific shRNA expressing MCF-7 cells were generated by transfecting with PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA) that contains puromycin resistance gene using FuGENE HD transfection reagent (Promega, Madison, WI, USA) according to the manufacturer’s instructions. ..
Plasmid Preparation:Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation
Article Snippet: In order to maintain the stable expression of PMCA4b transgenes and of the shRNA construct, 100 ng/ml puromycin dihydrochloride (sc-108071, Santa Cruz Biotechnology) was added to the culture media. .. PMCA4-specific shRNA MCF-7 cells were generated by using the PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA), as described previously ( Naffa et al ., 2021 ). ..
Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation.
Article Snippet: .. Generation of stable cell lines PMCA4-specific shRNA expressing MCF-7 cells were generated by transfecting with PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA) that contains puromycin resistance Fig. 8 | PMCA4b promotes central lumen formation in 3D cultures of MCF7 cells. a Phalloidin staining of mammospheres formed by parental, PMCA4specific shRNA (sh-PMCA4), GFP-PMCA4b and GFP-PMCA4bLA-expressing MCF-7 cell lines cultured inMatrigel for 10 days. b Schematic figures demonstrating structures of mammospheres based on results on (a). c Statistical comparisons of the ratios of mammospheres with central lumen in 3D cultures between parental, PMCA4-specific shRNA (sh-PMCA4), GFP-PMCA4b- and GFP-PMCA4bLAexpressing MCF-7 cell lines cultured in Matrigel. ..
Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation
Article Snippet: .. PMCA4-specific shRNA expressing MCF-7 cells were generated by transfecting with PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA) that contains puromycin resistance gene using FuGENE HD transfection reagent (Promega, Madison, WI, USA) according to the manufacturer’s instructions. ..
Stable Transfection:Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation.
Article Snippet: .. Generation of stable cell lines PMCA4-specific shRNA expressing MCF-7 cells were generated by transfecting with PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA) that contains puromycin resistance Fig. 8 | PMCA4b promotes central lumen formation in 3D cultures of MCF7 cells. a Phalloidin staining of mammospheres formed by parental, PMCA4specific shRNA (sh-PMCA4), GFP-PMCA4b and GFP-PMCA4bLA-expressing MCF-7 cell lines cultured inMatrigel for 10 days. b Schematic figures demonstrating structures of mammospheres based on results on (a). c Statistical comparisons of the ratios of mammospheres with central lumen in 3D cultures between parental, PMCA4-specific shRNA (sh-PMCA4), GFP-PMCA4b- and GFP-PMCA4bLAexpressing MCF-7 cell lines cultured in Matrigel. ..
Expressing:Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation.
Article Snippet: .. Generation of stable cell lines PMCA4-specific shRNA expressing MCF-7 cells were generated by transfecting with PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA) that contains puromycin resistance Fig. 8 | PMCA4b promotes central lumen formation in 3D cultures of MCF7 cells. a Phalloidin staining of mammospheres formed by parental, PMCA4specific shRNA (sh-PMCA4), GFP-PMCA4b and GFP-PMCA4bLA-expressing MCF-7 cell lines cultured inMatrigel for 10 days. b Schematic figures demonstrating structures of mammospheres based on results on (a). c Statistical comparisons of the ratios of mammospheres with central lumen in 3D cultures between parental, PMCA4-specific shRNA (sh-PMCA4), GFP-PMCA4b- and GFP-PMCA4bLAexpressing MCF-7 cell lines cultured in Matrigel. ..
Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation
Article Snippet: .. PMCA4-specific shRNA expressing MCF-7 cells were generated by transfecting with PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA) that contains puromycin resistance gene using FuGENE HD transfection reagent (Promega, Madison, WI, USA) according to the manufacturer’s instructions. ..
Staining:Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation.
Article Snippet: .. Generation of stable cell lines PMCA4-specific shRNA expressing MCF-7 cells were generated by transfecting with PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA) that contains puromycin resistance Fig. 8 | PMCA4b promotes central lumen formation in 3D cultures of MCF7 cells. a Phalloidin staining of mammospheres formed by parental, PMCA4specific shRNA (sh-PMCA4), GFP-PMCA4b and GFP-PMCA4bLA-expressing MCF-7 cell lines cultured inMatrigel for 10 days. b Schematic figures demonstrating structures of mammospheres based on results on (a). c Statistical comparisons of the ratios of mammospheres with central lumen in 3D cultures between parental, PMCA4-specific shRNA (sh-PMCA4), GFP-PMCA4b- and GFP-PMCA4bLAexpressing MCF-7 cell lines cultured in Matrigel. ..
Cell Culture:Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation.
Article Snippet: .. Generation of stable cell lines PMCA4-specific shRNA expressing MCF-7 cells were generated by transfecting with PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA) that contains puromycin resistance Fig. 8 | PMCA4b promotes central lumen formation in 3D cultures of MCF7 cells. a Phalloidin staining of mammospheres formed by parental, PMCA4specific shRNA (sh-PMCA4), GFP-PMCA4b and GFP-PMCA4bLA-expressing MCF-7 cell lines cultured inMatrigel for 10 days. b Schematic figures demonstrating structures of mammospheres based on results on (a). c Statistical comparisons of the ratios of mammospheres with central lumen in 3D cultures between parental, PMCA4-specific shRNA (sh-PMCA4), GFP-PMCA4b- and GFP-PMCA4bLAexpressing MCF-7 cell lines cultured in Matrigel. ..
Transfection:Article Title: The calcium pump PMCA4b promotes epithelial cell polarization and lumen formation
Article Snippet: .. PMCA4-specific shRNA expressing MCF-7 cells were generated by transfecting with PMCA4-specific shRNA plasmid (sc-42602-SH, Santa Cruz Biotechnology, Santa Cruz, CA, USA) that contains puromycin resistance gene using FuGENE HD transfection reagent (Promega, Madison, WI, USA) according to the manufacturer’s instructions. ..
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