Amplification:Article Title: Anti-cancer effects of curcumin on lung cancer through the inhibition of EZH2 and NOTCH1.
Article Snippet: .. We amplified the EZH2 coding sequence from pCMV6-EZH2 (Origene, Rockville, MD) using primer 1 (Supplementary Table S1) and cloned it into pcDNA3.1(+) (Invitrogen, Carlsbad, CA) between the NheI and KpnI sites to construct the vector expressing EZH2 (pcDNA3EZH2Flag). ezh2 promoter (–1772 to +112 relative to the start of the first exon) amplified with primer 2 (Supplementary Table S1) from genomic DNA of A549 cells or the randomly scrambled ezh2 promoter that was synthesized was inserted into the NheI and HindIII sites of pGL3-Basic (Promega, Madison, WI) to generate the ezh2 promoter luciferase reporter vector pGL3-EZH2P and the negative control vector pGL3-EZH2Pscram, respectively. .. To construct pMIR-EZH2 UTR, 263 bp of the EZH2 3′ UTR sequence was amplified and cloned into the luciferase reporter vector pMIR-REPORTTM Luciferase (Ambion/Life Technologies, Grand Island, NY) between the SpeI and HindIII sites as previously reported [18].
Sequencing:Article Title: Anti-cancer effects of curcumin on lung cancer through the inhibition of EZH2 and NOTCH1.
Article Snippet: .. We amplified the EZH2 coding sequence from pCMV6-EZH2 (Origene, Rockville, MD) using primer 1 (Supplementary Table S1) and cloned it into pcDNA3.1(+) (Invitrogen, Carlsbad, CA) between the NheI and KpnI sites to construct the vector expressing EZH2 (pcDNA3EZH2Flag). ezh2 promoter (–1772 to +112 relative to the start of the first exon) amplified with primer 2 (Supplementary Table S1) from genomic DNA of A549 cells or the randomly scrambled ezh2 promoter that was synthesized was inserted into the NheI and HindIII sites of pGL3-Basic (Promega, Madison, WI) to generate the ezh2 promoter luciferase reporter vector pGL3-EZH2P and the negative control vector pGL3-EZH2Pscram, respectively. .. To construct pMIR-EZH2 UTR, 263 bp of the EZH2 3′ UTR sequence was amplified and cloned into the luciferase reporter vector pMIR-REPORTTM Luciferase (Ambion/Life Technologies, Grand Island, NY) between the SpeI and HindIII sites as previously reported [18].
Clone Assay:Article Title: Anti-cancer effects of curcumin on lung cancer through the inhibition of EZH2 and NOTCH1.
Article Snippet: .. We amplified the EZH2 coding sequence from pCMV6-EZH2 (Origene, Rockville, MD) using primer 1 (Supplementary Table S1) and cloned it into pcDNA3.1(+) (Invitrogen, Carlsbad, CA) between the NheI and KpnI sites to construct the vector expressing EZH2 (pcDNA3EZH2Flag). ezh2 promoter (–1772 to +112 relative to the start of the first exon) amplified with primer 2 (Supplementary Table S1) from genomic DNA of A549 cells or the randomly scrambled ezh2 promoter that was synthesized was inserted into the NheI and HindIII sites of pGL3-Basic (Promega, Madison, WI) to generate the ezh2 promoter luciferase reporter vector pGL3-EZH2P and the negative control vector pGL3-EZH2Pscram, respectively. .. To construct pMIR-EZH2 UTR, 263 bp of the EZH2 3′ UTR sequence was amplified and cloned into the luciferase reporter vector pMIR-REPORTTM Luciferase (Ambion/Life Technologies, Grand Island, NY) between the SpeI and HindIII sites as previously reported [18].
Construct:Article Title: Anti-cancer effects of curcumin on lung cancer through the inhibition of EZH2 and NOTCH1.
Article Snippet: .. We amplified the EZH2 coding sequence from pCMV6-EZH2 (Origene, Rockville, MD) using primer 1 (Supplementary Table S1) and cloned it into pcDNA3.1(+) (Invitrogen, Carlsbad, CA) between the NheI and KpnI sites to construct the vector expressing EZH2 (pcDNA3EZH2Flag). ezh2 promoter (–1772 to +112 relative to the start of the first exon) amplified with primer 2 (Supplementary Table S1) from genomic DNA of A549 cells or the randomly scrambled ezh2 promoter that was synthesized was inserted into the NheI and HindIII sites of pGL3-Basic (Promega, Madison, WI) to generate the ezh2 promoter luciferase reporter vector pGL3-EZH2P and the negative control vector pGL3-EZH2Pscram, respectively. .. To construct pMIR-EZH2 UTR, 263 bp of the EZH2 3′ UTR sequence was amplified and cloned into the luciferase reporter vector pMIR-REPORTTM Luciferase (Ambion/Life Technologies, Grand Island, NY) between the SpeI and HindIII sites as previously reported [18].
Expressing:Article Title: Anti-cancer effects of curcumin on lung cancer through the inhibition of EZH2 and NOTCH1.
Article Snippet: .. We amplified the EZH2 coding sequence from pCMV6-EZH2 (Origene, Rockville, MD) using primer 1 (Supplementary Table S1) and cloned it into pcDNA3.1(+) (Invitrogen, Carlsbad, CA) between the NheI and KpnI sites to construct the vector expressing EZH2 (pcDNA3EZH2Flag). ezh2 promoter (–1772 to +112 relative to the start of the first exon) amplified with primer 2 (Supplementary Table S1) from genomic DNA of A549 cells or the randomly scrambled ezh2 promoter that was synthesized was inserted into the NheI and HindIII sites of pGL3-Basic (Promega, Madison, WI) to generate the ezh2 promoter luciferase reporter vector pGL3-EZH2P and the negative control vector pGL3-EZH2Pscram, respectively. .. To construct pMIR-EZH2 UTR, 263 bp of the EZH2 3′ UTR sequence was amplified and cloned into the luciferase reporter vector pMIR-REPORTTM Luciferase (Ambion/Life Technologies, Grand Island, NY) between the SpeI and HindIII sites as previously reported [18].
Synthesized:Article Title: Anti-cancer effects of curcumin on lung cancer through the inhibition of EZH2 and NOTCH1.
Article Snippet: .. We amplified the EZH2 coding sequence from pCMV6-EZH2 (Origene, Rockville, MD) using primer 1 (Supplementary Table S1) and cloned it into pcDNA3.1(+) (Invitrogen, Carlsbad, CA) between the NheI and KpnI sites to construct the vector expressing EZH2 (pcDNA3EZH2Flag). ezh2 promoter (–1772 to +112 relative to the start of the first exon) amplified with primer 2 (Supplementary Table S1) from genomic DNA of A549 cells or the randomly scrambled ezh2 promoter that was synthesized was inserted into the NheI and HindIII sites of pGL3-Basic (Promega, Madison, WI) to generate the ezh2 promoter luciferase reporter vector pGL3-EZH2P and the negative control vector pGL3-EZH2Pscram, respectively. .. To construct pMIR-EZH2 UTR, 263 bp of the EZH2 3′ UTR sequence was amplified and cloned into the luciferase reporter vector pMIR-REPORTTM Luciferase (Ambion/Life Technologies, Grand Island, NY) between the SpeI and HindIII sites as previously reported [18].
Luciferase:Article Title: Anti-cancer effects of curcumin on lung cancer through the inhibition of EZH2 and NOTCH1.
Article Snippet: .. We amplified the EZH2 coding sequence from pCMV6-EZH2 (Origene, Rockville, MD) using primer 1 (Supplementary Table S1) and cloned it into pcDNA3.1(+) (Invitrogen, Carlsbad, CA) between the NheI and KpnI sites to construct the vector expressing EZH2 (pcDNA3EZH2Flag). ezh2 promoter (–1772 to +112 relative to the start of the first exon) amplified with primer 2 (Supplementary Table S1) from genomic DNA of A549 cells or the randomly scrambled ezh2 promoter that was synthesized was inserted into the NheI and HindIII sites of pGL3-Basic (Promega, Madison, WI) to generate the ezh2 promoter luciferase reporter vector pGL3-EZH2P and the negative control vector pGL3-EZH2Pscram, respectively. .. To construct pMIR-EZH2 UTR, 263 bp of the EZH2 3′ UTR sequence was amplified and cloned into the luciferase reporter vector pMIR-REPORTTM Luciferase (Ambion/Life Technologies, Grand Island, NY) between the SpeI and HindIII sites as previously reported [18].
Negative Control:Article Title: Anti-cancer effects of curcumin on lung cancer through the inhibition of EZH2 and NOTCH1.
Article Snippet: .. We amplified the EZH2 coding sequence from pCMV6-EZH2 (Origene, Rockville, MD) using primer 1 (Supplementary Table S1) and cloned it into pcDNA3.1(+) (Invitrogen, Carlsbad, CA) between the NheI and KpnI sites to construct the vector expressing EZH2 (pcDNA3EZH2Flag). ezh2 promoter (–1772 to +112 relative to the start of the first exon) amplified with primer 2 (Supplementary Table S1) from genomic DNA of A549 cells or the randomly scrambled ezh2 promoter that was synthesized was inserted into the NheI and HindIII sites of pGL3-Basic (Promega, Madison, WI) to generate the ezh2 promoter luciferase reporter vector pGL3-EZH2P and the negative control vector pGL3-EZH2Pscram, respectively. .. To construct pMIR-EZH2 UTR, 263 bp of the EZH2 3′ UTR sequence was amplified and cloned into the luciferase reporter vector pMIR-REPORTTM Luciferase (Ambion/Life Technologies, Grand Island, NY) between the SpeI and HindIII sites as previously reported [18].
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