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Differential usage of LST1 uORF-containing exons during monocyte differentiation.

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posted on 09.05.2014, 03:21 authored by Christian Schiller, Carina Nowak, Kalliope N. Diakopoulos, Ulrich H. Weidle, Elisabeth H. Weiss

Cells from the monocytic cell lines U-937 and THP-1 were prompted to differentiate to macrophages by treatment with 80 nM TPA for 72 h. (A, B)LST1 transcripts containing either exon 1B or 1C were detected by PCR using specific primers and cDNA transcribed from total RNA which was isolated from treated and mock-treated cells. Amplificates for transcripts containing exon 1B and 1C were detected on an agarose gel at 90 and 125 bp, respectively. The amplified GAPDH sequence was detected at 393 bp. (C, D) U-937 and THP-1 cells were treated with TPA, RNA was isolated and cDNA was synthesised as described above. The amount of LST1 transcripts containing either exon 1B (LST1B) or exon 1C (LST1C) was assayed by quantitative PCR. LST1 transcript expression was normalised using GAPDH. A value of 100% was set for mock-treated cells. Mean values from 5 independent experiments are indicated within the columns +/− s.d. In both U-937 and THP-1 cells, treatment with TPA led to a distinct and significant (p = 0.009) increase in the number of transcripts containing exon 1C. For exon 1B containing splice variants only a modest, non-significant (p = 0.094) increase in transcript amounts after TPA treatment of both U-937 and THP-1 cells could be detected.