Figure 1: GNG12-AS1 is a stable lncRNA in the nucleus.
(a) Schematic representation of the GNG12-AS1 genomic locus (chr1: 68297971–68668670, hg19) relative to GNG12, DIRAS3 and WLS. The arrows show the direction of transcription. GNG12-AS1 exons are in green and are numbered as previously reported31. Probes used for FISH are marked. Intronic, labelled in red identify the site of transcription. Exonic, labelled in green, mark mature transcripts. (b) RNA distribution from the cytoplasm, nucleoplasm and chromatin in SUM159 cells as quantified by qRT–PCR. RPS18 and MALAT1 are positive controls for the cytoplasmic and chromatin fraction, respectively. Note enrichment of GNG12-AS1 in the chromatin fraction. Relative RNA levels are standardized to the geometric mean of GAPDH and β-actin. Error bars represent the s.e.m. values of three independent experiments. (c,d) Co-localization of exonic (green) and intronic (red) GNG12-AS1 probes in HB2 and SUM19 cells by single-molecule RNA FISH. GNG12-AS1 associates with its site of transcription (2.8% in SUM159, 6.8% in HB2; cis, c) but is also present at the other sites in the nucleus (1.7% in SUM159, 3.3% in HB2, trans, d). The numbers represent the percentage of cells positive for exonic and intronic FISH signal (n=132 for HB2; n=176 for SUM159). The nucleus was stained with 4,6-diamidino-2-phenylindole (DAPI). Scale bar, 7 μm.
