Identification of Host Kinase Genes Required for Influenza Virus Replication and the Regulatory Role of MicroRNAs
Figure 5
(A) A549 cells were transfected with 25 nM of miR-548d, -29a and -138 inhibitor/mimic for 48 hrs followed by RNA extraction and RT-qPCR with MAP3K1 specific primers. Expression data was normalized to 18S rRNA expression and shown as mean±SEM of independent experiments. *p<0.05 compared to control. (B) A549 cells mock/transfected with miR-548d inhibitor/mimic for 48 hrs were fixed with 4% formaldehyde in PBS and stained for MAPK1 protein using biotinylated rabbit anti-MAP3K1 antibody (Abcam ab69533) and detected with Streptavidin-Alexa488. Cells were analyzed by Arrayscan Cellomics VTI scanner and data analyzed by GraphPad Prism. (C) A549 cells mock/transfected with miR-548d inhibitor/mimic for 48 hrs were infected with A/WSN/33 (MOI = 0.001) for 48 hrs and stained for influenza NP protein using mouse-anti-NP coupled to Alexa-488 and analyzed as above in (B). Data show mean±SEM of two independent experiments. *p<0.05.