Generated:Article Title: Beclin 1 functions as a negative modulator of MLKL oligomerisation by integrating into the necrosome complex.
Article Snippet: .. We obtained and engineered pMSCVpuro-FLAG and pMSCVhygro-FLAG vectors from Addgene (#53178, #75083). pLKO.1 puro-shBECN1 and pLKO.1 hygro-shRIPK3 were generated using oligo annealing and cloning into an empty vector using following primers: shBECN1 #5F 5′-C CGGGATACCGACTTGTTCCTTACGCTCGAGCGTAA GGAACAAGTCGGTATCTTTTTG-3’, R 5′-AATTCAAA AAGATACCGACTTGTTCCTTACGCTCGAGCGTAAG GAACAAGTCGGTATC-3′, shBECN1 #7F 5′-CCGGCT AAGGAGCTGCCGTTATACTCTCGAGAGTATAACG GCAGCTCCTTAGTTTTTG-3′, R 5′-AATTCAAAAAC TAAGGAGCTGCCGTTATACTCTCGAGAGTATAAC GGCAGCTCCTTAG-3′, shRIPK3 F 5′-CCGGAACCAGC ACTCTCGTAATGATCTCGAGATCATTACGAGAGTG CTGGTTTTTTTG-3′, R 5′-AATTCAAAAAAACCAG CACTCTCGTAATGATCTCGAGATCATTACGAGAGT GCTGGTT-3′. shMLKL (TRCN0000196317) was purchased from Sigma-Aldrich. .. For transfection, the plasmids were incubated with polyethyleneimine (PEI) (Sigma-Aldrich) in serum-free media for 20min, and then added to 293T cells.
Article Title: Beclin 1 functions as a negative modulator of MLKL oligomerisation by integrating into the necrosome complex
Article Snippet: .. We obtained and engineered pMSCVpuro-FLAG and pMSCVhygro-FLAG vectors from Addgene (#53178, #75083). pLKO.1 puro-shBECN1 and pLKO.1 hygro-shRIPK3 were generated using oligo annealing and cloning into an empty vector using following primers: shBECN1 #5F 5′-CCGGGATACCGACTTGTTCCTTACGCTCGAGCGTAAGGAACAAGTCGGTATCTTTTTG-3’, R 5′-AATTCAAAAAGATACCGACTTGTTCCTTACGCTCGAGCGTAAGGAACAAGTCGGTATC-3′, shBECN1 #7F 5′-CCGGCTAAGGAGCTGCCGTTATACTCTCGAGAGTATAACGGCAGCTCCTTAGTTTTTG-3′, R 5′-AATTCAAAAACTAAGGAGCTGCCGTTATACTCTCGAGAGTATAACGGCAGCTCCTTAG-3′, shRIPK3 F 5′-CCGGAACCAGCACTCTCGTAATGATCTCGAGATCATTACGAGAGTGCTGGTTTTTTTG-3′, R 5′-AATTCAAAAAAACCAGCACTCTCGTAATGATCTCGAGATCATTACGAGAGTGCTGGTT-3′. shMLKL (TRCN0000196317) was purchased from Sigma-Aldrich. .. For transfection, the plasmids were incubated with polyethyleneimine (PEI) (Sigma-Aldrich) in serum-free media for 20 min, and then added to 293T cells.
Cloning:Article Title: Beclin 1 functions as a negative modulator of MLKL oligomerisation by integrating into the necrosome complex.
Article Snippet: .. We obtained and engineered pMSCVpuro-FLAG and pMSCVhygro-FLAG vectors from Addgene (#53178, #75083). pLKO.1 puro-shBECN1 and pLKO.1 hygro-shRIPK3 were generated using oligo annealing and cloning into an empty vector using following primers: shBECN1 #5F 5′-C CGGGATACCGACTTGTTCCTTACGCTCGAGCGTAA GGAACAAGTCGGTATCTTTTTG-3’, R 5′-AATTCAAA AAGATACCGACTTGTTCCTTACGCTCGAGCGTAAG GAACAAGTCGGTATC-3′, shBECN1 #7F 5′-CCGGCT AAGGAGCTGCCGTTATACTCTCGAGAGTATAACG GCAGCTCCTTAGTTTTTG-3′, R 5′-AATTCAAAAAC TAAGGAGCTGCCGTTATACTCTCGAGAGTATAAC GGCAGCTCCTTAG-3′, shRIPK3 F 5′-CCGGAACCAGC ACTCTCGTAATGATCTCGAGATCATTACGAGAGTG CTGGTTTTTTTG-3′, R 5′-AATTCAAAAAAACCAG CACTCTCGTAATGATCTCGAGATCATTACGAGAGT GCTGGTT-3′. shMLKL (TRCN0000196317) was purchased from Sigma-Aldrich. .. For transfection, the plasmids were incubated with polyethyleneimine (PEI) (Sigma-Aldrich) in serum-free media for 20min, and then added to 293T cells.
Article Title: Beclin 1 functions as a negative modulator of MLKL oligomerisation by integrating into the necrosome complex
Article Snippet: .. We obtained and engineered pMSCVpuro-FLAG and pMSCVhygro-FLAG vectors from Addgene (#53178, #75083). pLKO.1 puro-shBECN1 and pLKO.1 hygro-shRIPK3 were generated using oligo annealing and cloning into an empty vector using following primers: shBECN1 #5F 5′-CCGGGATACCGACTTGTTCCTTACGCTCGAGCGTAAGGAACAAGTCGGTATCTTTTTG-3’, R 5′-AATTCAAAAAGATACCGACTTGTTCCTTACGCTCGAGCGTAAGGAACAAGTCGGTATC-3′, shBECN1 #7F 5′-CCGGCTAAGGAGCTGCCGTTATACTCTCGAGAGTATAACGGCAGCTCCTTAGTTTTTG-3′, R 5′-AATTCAAAAACTAAGGAGCTGCCGTTATACTCTCGAGAGTATAACGGCAGCTCCTTAG-3′, shRIPK3 F 5′-CCGGAACCAGCACTCTCGTAATGATCTCGAGATCATTACGAGAGTGCTGGTTTTTTTG-3′, R 5′-AATTCAAAAAAACCAGCACTCTCGTAATGATCTCGAGATCATTACGAGAGTGCTGGTT-3′. shMLKL (TRCN0000196317) was purchased from Sigma-Aldrich. .. For transfection, the plasmids were incubated with polyethyleneimine (PEI) (Sigma-Aldrich) in serum-free media for 20 min, and then added to 293T cells.
Plasmid Preparation:Article Title: Beclin 1 functions as a negative modulator of MLKL oligomerisation by integrating into the necrosome complex.
Article Snippet: .. We obtained and engineered pMSCVpuro-FLAG and pMSCVhygro-FLAG vectors from Addgene (#53178, #75083). pLKO.1 puro-shBECN1 and pLKO.1 hygro-shRIPK3 were generated using oligo annealing and cloning into an empty vector using following primers: shBECN1 #5F 5′-C CGGGATACCGACTTGTTCCTTACGCTCGAGCGTAA GGAACAAGTCGGTATCTTTTTG-3’, R 5′-AATTCAAA AAGATACCGACTTGTTCCTTACGCTCGAGCGTAAG GAACAAGTCGGTATC-3′, shBECN1 #7F 5′-CCGGCT AAGGAGCTGCCGTTATACTCTCGAGAGTATAACG GCAGCTCCTTAGTTTTTG-3′, R 5′-AATTCAAAAAC TAAGGAGCTGCCGTTATACTCTCGAGAGTATAAC GGCAGCTCCTTAG-3′, shRIPK3 F 5′-CCGGAACCAGC ACTCTCGTAATGATCTCGAGATCATTACGAGAGTG CTGGTTTTTTTG-3′, R 5′-AATTCAAAAAAACCAG CACTCTCGTAATGATCTCGAGATCATTACGAGAGT GCTGGTT-3′. shMLKL (TRCN0000196317) was purchased from Sigma-Aldrich. .. For transfection, the plasmids were incubated with polyethyleneimine (PEI) (Sigma-Aldrich) in serum-free media for 20min, and then added to 293T cells.
Article Title: Beclin 1 functions as a negative modulator of MLKL oligomerisation by integrating into the necrosome complex
Article Snippet: .. We obtained and engineered pMSCVpuro-FLAG and pMSCVhygro-FLAG vectors from Addgene (#53178, #75083). pLKO.1 puro-shBECN1 and pLKO.1 hygro-shRIPK3 were generated using oligo annealing and cloning into an empty vector using following primers: shBECN1 #5F 5′-CCGGGATACCGACTTGTTCCTTACGCTCGAGCGTAAGGAACAAGTCGGTATCTTTTTG-3’, R 5′-AATTCAAAAAGATACCGACTTGTTCCTTACGCTCGAGCGTAAGGAACAAGTCGGTATC-3′, shBECN1 #7F 5′-CCGGCTAAGGAGCTGCCGTTATACTCTCGAGAGTATAACGGCAGCTCCTTAGTTTTTG-3′, R 5′-AATTCAAAAACTAAGGAGCTGCCGTTATACTCTCGAGAGTATAACGGCAGCTCCTTAG-3′, shRIPK3 F 5′-CCGGAACCAGCACTCTCGTAATGATCTCGAGATCATTACGAGAGTGCTGGTTTTTTTG-3′, R 5′-AATTCAAAAAAACCAGCACTCTCGTAATGATCTCGAGATCATTACGAGAGTGCTGGTT-3′. shMLKL (TRCN0000196317) was purchased from Sigma-Aldrich. .. For transfection, the plasmids were incubated with polyethyleneimine (PEI) (Sigma-Aldrich) in serum-free media for 20 min, and then added to 293T cells.
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