Fig. 1.

Introduction of siRNA duplexes by nucleofection and lipofectamine-based transfection results in efficient knockdown of galectin-3 in filter-grown Madin-Darby canine kidney (MDCK) cells. A: cells were nucleofected with the indicated siRNAs and plated onto filters the following day. Five days after nucleofection, cells were solubilized and lysates were analyzed by Western blotting to detect galectin-3 or β-actin (as a loading control). Untransfected cells (UNTRANS) plated under identical conditions were included as an additional control. B: MDCK cells suspended in MEM were incubated with siRNA duplexes and lipofectamine in the apical chamber of Transwell filter cups. Cells were cultured for 4 days before solubilization and Western blotting. Knockdown efficiency was typically >85% in samples nucleofected with galectin-3 siRNA and slightly lower (∼80%) in lipofectamine-treated cells. CTRL, control.