Fig. 1: Characterization of Mammary Tumor Associated RNA 25 (MaTAR25).

a Representation of the MaTAR25 gene locus. MaTAR25 is an intergenic lncRNA gene located on mouse chromosome 2, and the MaTAR25 RNA transcript contains 2 exons and a poly (A) tail. b 5′ and 3′ rapid amplification of cDNA ends (RACE) was performed to identify the full-length MaTAR25 transcript. Three independent experiments were performed, and a representative gel image is shown. c The full-length MaTAR25 transcript was confirmed by northern blot analysis to be ~2000 nt. In total, 20 μg or 30 μg of total RNA samples extracted from MMTV-PyMT primary cells were electrophoresed on a 1% agarose gel and probed. Two independent experiments were performed, and a representative gel image is shown. d In vitro transcription and translation reactions were performed to confirm that MaTAR25 does not produce a peptide. The reaction products were loaded on a 4–20% gradient SDS-PAGE gel, and the signals were detected by HRP-conjugated streptavidin. Luciferase control DNA and Xenopus laevis Histone H2B (HISTH2B) expressing plasmids were used as positive controls and empty vector as a negative control. Three independent experiments were performed, and a representative gel image is shown. e Representative smRNA-FISH images showing localization of MaTAR25 RNA transcripts (red) within nuclei of MMTV-PyMT and MMTV-Neu-NDL primary cells. Scale bars are 10 μm.