VIPR2 Expression and Modulation

The pCMV6-AN-Myc-DDK vector (PS100016) and a negative control siRNA (S10C-0600) were purchased from Cosmo Bio (Tokyo, Japan). The pEGFP-N2 vector and pmCherry-N1 vector were purchased from Takara Clontech (Kyoto, Japan). The pCMV6-VIPR2-Myc-DDK plasmid was constructed from the pCMV6-AN-Myc-DDK vector and VIPR2 cDNA. The VIPR2-Myc region from the resultant plasmid was cloned into the pEGFP-N2 vector, and the VIPR2-Myc-DDK region was cloned into pmCherry-N1 vector. The AKT-PH-EGFP vector was described previously (34 (link)). Human VIPR2-siRNAs (si1, 3024813653-000080 and -000090; si2, 3024813653-000020 and -000030; si3, 3024813653-000050 and -000060) were purchased from Sigma-Aldrich (St. Louis, MO, USA). A VIPR2-selective antagonistic peptide KS-133 was synthesized at SCRUM Inc. (Tokyo, Japan) as previously reported (35 (link)).