Figure 2
From: The chicken miR-150 targets the avian orthologue of the functional zebrafish MYB 3'UTR target site
gga-miR-150 ectopically produced in stable lymphoid chicken PA9 and MSB-1 cells represses the avian MYB reporter. (A) gga-miR-150 and gga-miR-150mut detected by northern blotting. Positive control: spleen leukocytes from a 4-week-old chicken. Northern blots were performed with a 32P-5' end-labelled DNA oligonucleotide probe complementary to the miRNA. (B) The MYB reporter was repressed in PA9 or MSB-1 cells producing gga-miR-150. PA9 and MSB-1 cells were electroporated with 40 Ī¼g of avian MYB WT pRL-TK reporter and 500 ng of the normalising plasmid containing the firefly luciferase gene, pcDNAMLuc. Luciferase activity was determined 24 hours after transfection. Renilla luciferase activity was normalised with respect to firefly luciferase activity and luciferase activity in PA9 and MSB-1 cells producing gga-miR-150mut was set at 100%. The data from one experiment representative of the three carried out are shown, with all three assays generating essentially identical results (* significant repression, P < 0.05, Student's t-test comparisons with PA9T or MSB-1T150mut).