Pegfp c1 pkm2

The vectors pWZL Neo Myr Flag PKM2 and pEGFP-C1-PKM2 were purchased from Addgene (plasmid #20585 and plasmid #64698, deposited by Jean Zhao and Axel Ullrich, respectively). The Glut1 promoter reporter clone (pEZXPG02.1-Glut1) was purchased from Genecopoeia (USA). The pTIG1-myc vector has been described previously (Wu et al., 2011 (link)). The DNAJC8 cDNA fragment was amplified from pDNAJC8/PACT2 using 5′ (5′-TGGCTAGCATGGCGGCTTCA GGAGAGAGC-3′) and 3′ (5′-GACTCGAGCTCACGTTGCTCC ATTTTTACTTTCGG-3′) primers and then was subcloned inframe into the NheI-XhoI sites of PCR3.1-Flag (Dr. Yi-Ling Lin, Institute of Biomedical Science, Academia Sinica, Taiwan) to generate pDNAJC8-Flag. The DNAJC8 cDNA fragment was amplified from pDNAJC8-Flag using 5′ (5′-TCGAATTCTATGG CGGCTTCAGGAG-3′) and 3′ (5′-GTGGATCCCTCACGTTGCT CCATTTTTA-3′) primers and then subcloned in-frame into the EcoRI-BamHI sites of the pEGFP-C1 vector (Clontech Laboratories, USA) to generate pEGFP-DNAJC8. The PKM2 cDNA fragment was amplified from pWZL Neo Myr Flag PKM2 using 5′ (5′-TCGAATTCGATGTCGAAGCCCCATAGTG-3′) and 3′ (5′-GTGGATCC CGGCACAGGAACAACACGC-3′) primers and then subcloned in-frame into the EcoRI-BamHI sites of the pcDNA-myc-His vector (Invitrogen, USA) to generate pPKM2-myc.

Plasmid pRP-Puro-CMV-CC16 and its vector pRP-Puro-CMV were purchased from VectorBuilder (Chicago, IL). Human HSP60 cDNA amplified by RT-PCR was subcloned into the Kpn I and Xho I sites of pcDNA3.1 vector (CMV promoter; Invitrogen) to obtain pcDNA-HSP60, and the primer sequences used for cloning are provided in Table S3. pEGFP-C1-PKM2 (Addgene plasmid #64698) was a gift from Axel Ullrich.⁷³ (link)