Efficient siRNA Delivery by the Cationic Liposome DOTAP in Human Hematopoietic Stem Cells Differentiating into Dendritic Cells
Silencing of cathepsin S during HSC differentiation. (a) Levels of cathepsin S were assessed by Western blotting in dendritic cells generated from the differentiation in vitro of HSCs. During the 14-day differentiation process, cells were subjected to DOTAP-based transfection rounds on day 0, 3, and 9 using a CTSS-targeting siRNA as described in the Materials and Methods. Untreated and scrambled siRNA-transfected cells were employed for control purposes. Levels of precursor (66 KDa) and mature (26 KDa) forms of cathepsin S at different time points (i.e., days 7 and 14) are indicated. -actin was used to normalize the signals. (b) Shown is the same analysis as in panel A, except for the number of transfections, that were performed on day 0 and 3 only. Graphs are representative of experiments consistently reproduced using at least 20 different populations of HSCs.
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