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. 2008 Aug 25;28(21):6609–6619. doi: 10.1128/MCB.00398-08

FIG. 3.

FIG. 3.

Transcriptional unit of miR302-367 gene. (A) 5′ RACE assay shows specific PCR amplification only in the presence of RT and TAP treatment (lane 2). Identical reactions without TAP treatment (lane 3) or without RT (lane 4) gave no amplification. M, molecular marker. (B) Sequence diagram depicting the gene structure and main genomic components of the human miR302-367 transcriptional unit. The first nucleotide to be transcribed is numbered as +1 (transcription start). Canonical sequence motifs for TATA box and polyadenylation signal are boxed. Intron sequences are shown in brackets and shortened due to their extension. The miR302-367 cluster is located within the first intron. (C) Adapted 3′ RACE assay displaying specific PCR product. No band was observed in the absence of RT. Two bands illustrated by arrows were clearly detectable. (D) Detection of putative polyadenylated unspliced pri-miRNA by Northern blot analysis in NTERA-2. Total RNA (7 μg) was loaded in lane 1. From the same amount of total RNA, poly(A)− (lane 2) and poly(A)+ (lane 3) fractions were loaded.