The replication of HCMV was suppressed by miR-221. (A) NPCs were transfected with mimics NC or miR-221 mimics for 24 h. The miR-221 expression level was checked by qRT-PCR. (B-C) NPCs were treated with mimics NC or miR-221 mimics for 24 h, followed by HCMV infection for 24 h. The HCMV copy numbers were checked by qRT-PCR assay at indicated times postinfection (B). HCMV titer was evaluated by plaque assay at indicated times post infection (C). (D) NPCs were treated with inhibitor NC or miR-221 inhibitor for 24 h, followed by HCMV infection for 24 h. The miR-221 expression level was checked by qRT-PCR. (E-F) NPCs were treated with inhibitor NC or miR-221 inhibitor for 24 h, followed by HCMV infection for 24 h. The HCMV copy numbers were checked by qRT-PCR assay at indicated times postinfection(E). HCMV titer was evaluated by plaque assay at indicated times post infection (F). (G-H) NPCs were treated with mimics NC, miR-221 mimics or inhibitor NC, miR-221 inhibitor for 24 h, followed by HCMV infection for 24 h. The expression levels of HCMV viral proteins IE2 and PP65 were checked by immunoblot (G). The expression levels of HCMV viral proteins IE1 was detected by immunofluorescence staining (H). All these experiments have been repeated three times, and the representative data were shown. (* p < 0.05, ** p < 0.01 vs mimics NC or inhibitor NC group).