PLGA-Mediated siRNA Delivery for Cyclin B1 Knockdown

Poly(d,l-lactic-co-glycolic acid) (PLGA7510; M_W, 10,000) was purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Human cyclin B1-specific siRNA, which was modified with a thiol group at the 3ʹ end of the sense or antisense strand, was purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). The siRNA sequences were as follows: cyclin B1 siRNA sense: 5ʹ-GGC GAA GAU CAA CAU GGC ATT-3ʹ; and cyclin B1 siRNA antisense: 5ʹ-UGC CAU GUU GAU CUU CGC CTT-3ʹ^{25 (link)}. Negative control siRNA was also purchased from Thermo Fisher Scientific, Inc. (Waltham). Fetal calf serum (FCS) and Roswell Park Memorial Institute (RPMI) 1640 were purchased from Gibco (Cergy-Pontoise, France). Anti-human CD71 monoclonal antibody, phycoerythrin (PE) anti-human CD71 antibody (Clone: OKT9) and were purchased from Thermo Fisher Scientific Inc. Normal mice IgG1 kappa monoclonal, which was used as the control IgG, was purchased from Abcam (ab91353, Cambridge, UK). A Pierce IgG F(abʹ)2 preparation kit was purchased from Thermo Fisher Scientific Inc. Cell Counting Kit-8 (CCK-8) reagent was purchased from Dojindo Laboratories (Tokyo, Japan). Alexa488-labeled siRNA was purchased from Thermo Fisher Scientific, Inc. (Waltham). Cy5-labeled siRNA was purchased from Nippon Gene (Toyama, Japan). FITC-labeled PLGA was synthesized by the NARD Institute, Ltd. (Amagasaki, Japan).