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PLoS Genet. 13. DUX4-induced dsRNA and MYC mRNA stabilization activate apoptotic pathways in human cell models of facioscapulohumeral dystrophy. 2018

Shadle, S.C., Zhong, J.W., Campbell, A.E., Conerly, M.L., Jagannathan, S., Wong, C.J., Morello, T.D., van der Maarel, S.M., Tapscott, S.J.

Notes: DUX4 is dysregulated in Facioscapulohumeral dystrophy (FSHD) leading to cellular apoptosis and disease. The genes and pathways involved in DUX4-mediated apoptosis are uncharacterized. Here, an siRNA screen to identify involved components is performed, and cell survival was measured using CellTiter-Glo. A counter screen to identify false positives which inhibited DUX4 induction by doxycycline was done using the ONE-Glo EX Luciferase Assay System. (5135)

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Cancer Res. 78. Nuclear IGF1R Interacts with regulatory regions of chromatin to promote RNA POlymerase II recruitment and gene expression associated with advanced tumor stage. 2018

Aleksic, T., Gray, N., Wu, X., Rieunier, G., Osher, E., Mills, J., Verrill, C., Bryant, R.J., Han, C., Hutchinson, K., Lambert, A.G., Kumar, R., Hamdy, F.C., Weyer-Czernilofsky, U., Sanderson, M.P., Bogenrieder, T., Taylor, S., Macaulay, V.M.

Notes: IGF-1R is internalized and localized to the nucleus in malignant prostate cancers. Here, the role of IGF-1R internalization is investigated using ChIP-seq. IGF-1R is shown to bind near transcription start sites of JUN and FAM21 genes and col-localized with RNA polymerase II. The proximal JUN promoter was cloned upstream of a luciferase reporter and luciferase activity was measured with the ONE-Glo EX Luciferase Assay. The presence of IDF-1R significantly increased luminescence. (5134)

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Biochem. Pharmacol. 136, 62–75. Real-time analysis of the binding of fluorescent VEGF165a to VEGFR2 in living cells: Effect of receptor tyrosine kinase inhibitors and fate of internalized agonist-receptor complexes. 2017

Kilpatrick, L. E., Friedman-Ohana, R., Alcobia, D. C., Riching, K., Peach, C. J., Wheal, A. J., Briddon, S. J., Robers, M. B., Zimmerman, K., Machleidt, T., Wood, K. V., Woolard, J. and Hill, S. J.

Notes: Vascular endothelial growth factor (VEGF) receptor interactions are observed using the NanoBRET™ Protein-Protein Interaction System. A novel method of labeling VEGF at a single N-terminal cysteine (TMR) to maintain full activity is presented. NanoLuc®-VEGFR2 and VEGF-TMR are used in conjunction to monitor an interaction and internalization into intracellular endosomes. The effect of receptor tyrosine kinase inhibitors such as Cediranib and vandetanib on internalization in living cells is assessed. (5060)

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