Figure 2. siRNAs Electroporated into hMSC and NHDF-neo Cells. Six siRNAs (1.5 g) targeting CDK2, JAK1, p53, GAPDH, uPAR, PKR, as well as a negative control siRNA, were electroporated into hMSC cells (A). Seven siRNAs (1.5 g) targeting Cyclin D1, NFκBp65, STAT1, PKCα, JAK1, RAF1, MMP-2, and a negative control siRNA were individually electroporated into NHDF-neo cells (B). All electroporations were conducted using the siPORT siRNA Electroporation Kit. 48 hours after electroporation, the cells were harvested and analyzed by real-time RT-PCR for corresponding gene expression levels. 18S rRNA levels were used to normalize the expression data. Percent remaining gene expression was calculated as a percentage of target mRNA compared with target mRNA from cells electroporated with the negative control siRNA.
Effective Gene Silencing in Primary and Difficult-to-Transfect Cells
In Figure 3, several different cell types were electroporated
with the GAPDH control siRNA using individually optimized electroporation
conditions. 48 hours after electroporation, GAPDH mRNA levels were reduced
by 70% or more in each cell type. Cell viability of 70% or greater was