pCMV‐CDK6 and pEGFP‐histone‐H1 vectors were described (Baldassarre et al, 1999 , 2000 ; Sonego et al, 2013 ). pFLAG‐FOXO3 was a gift from K. Guan (#13507 Addgene Inc.) and was used to generate the GFP‐FOXO3 vectors, using the pEGFP‐C1 vector (Clontech). CDK6 and FOXO3 point mutants were generated using the QuikChange Site‐Directed Mutagenesis Kit (Agilent). GST‐FOXO3 deletion mutants were generated by cloning FOXO3 PCR fragments in the pGEX vector. pLKO shRNAs (Sigma) and primers are listed in Appendix Table S3 .
Plko shrnas
Manufactured by Merck Group
PLKO shRNAs are a type of laboratory equipment used for gene silencing experiments. They contain short hairpin RNA (shRNA) sequences that can be used to knockdown the expression of target genes in cell lines and model organisms. The core function of PLKO shRNAs is to provide a tool for researchers to study gene function and cellular pathways.
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Lab products found in correlation
2 protocols using plko shrnas
1
Plasmid Vectors for Cell Studies
2
Generating Mutant SGK2 Plasmids
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pDONR223 SGK2 was a gift from William Hahn & David Root (Addgene plasmid #23378) and it was used to generate the pEGFP SGK2 vector, using pEGFP-C1 vector (Clontech), and pLPC SGK2 vector, using the pLPC vector gently provided by Dr. R. Maestro (CRO Aviano). Site-directed mutagenesis was used to generate SGK2 DN (SGK2 T193A/S356A) mutant, SGK2 CA (SGK2 S356D) mutant and SGK2 Kinase Dead (KD) (SGK2 K64M) (primers listed in Table S1 ) with QuikChange Site-Directed Mutagenesis Kit (Agilent). mRFP-EGFP-LC3 was a gift from Tamotsu Yoshimori (Addgene plasmid # 21074). pLKO shRNAs were purchased from Sigma-Aldrich Co and they are listed in Table S1 .
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