Figure 6.

An in vitro protocol rapidly tests the efficiency of multiple candidate miRNAs against Axonin1. COS-7 cells were repeatedly transfected with pRFPRNAi vectors expressing different miRNAs (as indicated). In each case, the expression of RFP showed that the majority of cells were transfected (A, representative image). Cells were then co-transfected with a construct driving the expression of myc-tagged Axonin1. Immunolabeling for myc 24 h later using the 9E10 antibody (B–I) revealed the ability of the different miRNAs to suppress Axonin1 expression relative to control conditions. The images are from one representative repeat of this experiment. Scale bar: 200 µm.