MISSION^® shRNA Lentiviral Transduction

Figure 1.Five individual clones from MISSION^® shRNA Target Set NM_002467 were co-transfected with a lentivirus packaging plasmid into HEK 293T cells. The resulting lentiviral particles were used to infect the human lung cancer cell line, H1299. 24 hours post-infection, 2.5 mg/mL of puromycin was added to select for infected cells. Infection efficiency was approximately 95%. 72 hours post-selection, cells were harvested. C-MYC message RNA level was detected by real-time PCR. Actin mRNA was used as an internal control.

Figure 2.Protein lysates from the shRNA infected cells were analyzed by western blotting using a c-MYC antibody. C-MYC protein levels corresponds to the mRNA knockdown shown in Figure 1.