Downregulation of the Host Gene jigr1 by miR-92 Is Essential for Neuroblast Self-Renewal in Drosophila (original) (raw)
Fig 3
miR-92a and miR-92b are transcribed in the same transcriptional unit.
(A) Schematic representation of the genomic arrangement of miR-92a, miR-92b and jigr1 based on results from this study. Red lines indicate deleted regions. (B) Northern blot analysis of total RNA extracted from third instar larval heads of the indicated genotypes. RNA was probed for miR-92a, miR-92b, U6, Jigr1 (exon 6 probe), Jigr1 (long 3’UTR probe) and rp49. Deletion lines, Del #4 and Del #7, are homozygous lethal but in trans to the bigger deficiency covering the locus, Def(3R)BSC321, they are viable and used like this for the analysis. (C) Northern blot analysis of RNA from wild type whole third instar larvae (lane 1), wild type larval heads (lane 2) and Del #1 (lane 3) third instar larval heads. Probes specific for all the isoforms of jigr1 (exon 6 probe, first panel), for the long 3’UTR (second panel), and for alternatively spliced noncoding exon 2 (third panel), and rp49 (bottom panel) were used. (D) Schematic representation of jigr1 constructs. (E) Northern blot analysis of total RNA from HEK 293T cells transfected with UAST-_jigr1_-long-3’UTR or UAST-_jigr1_-short-3’UTR plasmids together with the Actin-Gal4 plasmid. Cells transfected with UAST-_jigr1_-long-3’UTR alone served as negative controls. miR-92b, miR-92a, and control U6 probes were used. The miR-92b probe recognizes endogenous miR-92b in HEK 293T cells.