FIG 1.

Lentiviral copy number increases with decreasing amount of LDLR-encoding plasmid cotransfected with the virus-producing plasmids used in the positive control. (A) Viral RNA in the supernatant of HEK293 cells was determined by RT-qPCR. The negative controls (NegC) were psPAX2 and pMD2.G, and the positive controls (PosC) were pHR-CMV-EGFP, psPAX2, and pMD2.G. Data are the arithmetic means of 3 biological replicates ± standard errors of the means (SEM). The presence of LDLR decreases the copy number significantly [one-way analysis of variance (ANOVA), F(4, 2) = 356.18], and the copy number is inversely related to the concentration of LDLR [one-way ANOVA, F(3, 2) = 373.33]. Wedges indicate the amount of the LDLR-encoding plasmid relative to pHR-CMV-EGFP used in the transfection (LDLR, 1:2 to 1:16). ***, P < 0.0005. (B) Western blot analysis of tagged LDLR present in lysates of the transfected HEK293 cells from panel A. Scanned film was automatically white corrected using GIMP software. (C) Correlation between relative LDLR expression (data derived from panel B via the GelAnalyzer plugin of ImageJ software) and copy number (data from panel A). Note the very high correlation of R² = 0.9881.