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. 2009 Mar 17;6(3):763–771. doi: 10.1021/mp900023v

Figure 2.

Figure 2

Inhibition of clathrin-mediated endocytosis significantly blocks uptake of siRNA lipoplexes but not functional siRNA delivery. BSC cells were treated with chlorpromazine (Chlorp, 10 μg/mL) or cytochalasin D (CytoD, 100 μM) for 1 h followed by siRNA lipoplex transfection for 4 h in the presence of the inhibitors. After washing, siRNA lipoplex uptake was measured by flow cytometry (see for details). Transfected cells were cultured for another 20 h before CyB transcript level was measured by real-time PCR. (A) Uptake of Cy5-siRNA lipoplexes in the absence or presence of inhibitors. (B) Relative levels of CyB transcript in the presence or absence of inhibitors as normalized to the level of untreated cells. (C) Relative levels of CyB transcript in BSC cells that expressed the wild-type (WT) or K44A dynamin. BSC cells that stably express either the WT or K44A dynamin were transfected with siRNA lipoplexes for 4 h, washed, cultured for 20 h, and then assayed for CyB transcript level. Data shown are mean ± SD (n = 4). Representative data from one of three to five experiments are shown.