Journal: Oncogene

Article Title: Alternative splicing of the imprinted candidate tumor suppressor gene ZAC regulates its antiproliferative and DNA binding activities.

doi: 10.1038/sj.onc.1204237

Figure Lengend Snippet: Figure 1 Identi®cation of an alternatively spliced variant of ZAC. (a) Genomic organization of ZAC 3'UTR, coding region and proximal 5'UTR. ZAC exons were numbered from the last one (exon n). The coding region (black boxes) was entirely comprised in two exons. ZF1 and most of ZF2 were encoded by exon n-1 whereas ZF3-7 were encoded by the last exon. (b) Sequences of exon-intron boundaries for exons n, n-1 and n-2 (c) RT ± PCR on RNA prepared from various human tissues. 2.5 ng polyA+ RNA (pituitary gland) and 100 ng total RNA (placenta and mammary gland, ovary, kidney, adrenal gland and fetal liver) were reverse transcribed and ampli®ed by PCR with primers located in exons n-2 and n. Numbers denote RNA samples prepared from dierent individuals. Equal volume of each RT ± PCR reaction was electrophoresed through a 1% agarose gel, blotted and probed with a probe recognizing both ZAC variants. Primers speci®c for b-actin were used to control for equal RNA input. Linear ampli®cation of both variants was veri®ed by varying the ratio of plasmids comprising ZAC and ZACD2 (right panel)

Article Snippet: Pituitary gland poly A+ RNA, human fetal liver and human kidney total RNA were from Clontech (Palo Alto, CA, USA) and human ovary total RNA was from Research Genetics, Inc. (Huntsville, AL, USA).

Techniques: Variant Assay, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription, Agarose Gel Electrophoresis, Control