TransIT-siQUEST® Transfection Reagent

Figure 1. Inhibition of PPAR-alpha Expression in Primary Mouse Hepatocytes Using the TransIT-siQUEST® Reagent. Primary mouse hepatocytes were transfected with an anti-PPAR-alpha siRNA or a non-targeting control siRNA using the TransIT-siQUEST® Reagent. Twenty-four hours post-transfection, the amount of PPAR-alpha mRNA was measured relative to GAPDH mRNA levels using qRT-PCR and then scaled to the expression level of the specific target mRNA in the Opti-MEM® media only (untreated) control.

Figure 2. Inhibition of Stably Expressed Firefly Luciferase Using the TransIT-siQUEST® Reagent. Cell lines stably expressing firefly luciferase were transfected with an anti-firefly luciferase siRNA or a reagent alone control using the TransIT-siQUEST® Reagent. Twenty-four hours post-transfection, luciferase expression was measured and compared to the reagent alone control.

Figure 3. Efficient Target Gene Knockdown in Selected Cell Lines Using TransIT-siQUEST® Reagent. Reporter plasmids expressing both firefly and sea pansy luciferase were co-transfected into the indicated cell lines using TransIT® Plasmid Transfection Reagents. Targeted knockdown was achieved by transfection of an anti-firefly luciferase siRNA using the TransIT-siQUEST® Reagent. Twenty-four hours later, firefly luciferase expression was normalized to sea pansy luciferase expression and compared to the reagent alone control.

Figure 4. Visualization of Fluorescently-Labeled siRNA Transfected with the TransIT-siQUEST® Reagent. (A) HeLa cells were transfected with a Cy™3-labeled siRNA using the TransIT-siQUEST® Reagent or (B) a reagent alone control. At 4 hours post-transfection, the cells were fixed, counterstained, and analyzed by confocal microscopy to visualize the siRNA (red), nuclei (blue), and actin (green).