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Figure 1 | Epigenetics & Chromatin

Figure 1

From: Knockdown of menin affects pre-mRNA processing and promoter fidelity at the interferon-gamma inducible IRF1 gene

Figure 1

Characterization of MEN1-shRNAmir and non-silencing shRNAmir cell lines. (a) Graphic depiction of the pTRIPZ shRNAmir vector. The pTRIPZ transactivator, reverse tetracycline transactivator 3 (rtTA3), binds to and activates expression from a TRE promoter in the presence of doxycycline, for the inducible expression of an shRNAmir. (b) Western blotting of whole cell extracts from a stable 2fTGH cell line either expressing the inducible knockdown shRNAmir vector targeting MEN1 (shMEN1) or a nonsilencing shRNAmir (NS) vector were subjected to SDS-PAGE, blotted on nitrocellulose and a MEN1 antibody was used to develop the blots. STAT1 served as a loading control. Quantification was performed using Image J. (c) Graphic depiction of the IRF1 gene locus (approximately 9 kb) indicating the TSS, location of the two polyadenylation signals (inverted triangles) and locations of ChIP qPCR primers (dashes) and RACE primers (arrows) used in this study. * indicates primers also used in qRT-PCR. ChIP: chromatin immunoprecipitation; kb: kilobases; MEN1: multiple endocrine neoplasia type 1; qPCR: quantitative PCR; qRT-PCR: quantitative real time PCR; RACE: rapid amplification of cDNA ends; STAT1: signal transducer and activator of transcription 1; TSS: transcription start site.

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