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. 2011 Oct 5;6(10):e25924. doi: 10.1371/journal.pone.0025924

Figure 1. Electrostatic interaction between CADY and HSPGs is not required for efficient transfection.

Figure 1

(A) HeLa cells were pre-treated for 30 min with free medium or medium containing 5 µg/ml heparin prior to addition of 80 nM siRNA complexed to CADY at a 1∶40 molar ratio. Cells were incubated in the presence of heparin for 1 hr, then extensively washed in PBS and replaced in DMEN containing 10% FCS. Cells were harvested after 48 hours and protein levels were analysed by Western blotting. CHO wild type (WT) (B), glycosaminoglycan deficient (gl−/−) (C) and heparan sulfate deficient (HS−/−) (D) cells were transfected with 80 nM FITC-labeled siRNA complexed to CADY at a 1∶20 molar ratio. After 1.5 hrs, cells were extensively washed, trypsinized and analyzed by FACS.