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Yes‐associated protein 1 (YAP) and transcriptional coactivator with PDZ‐binding motif (TAZ) are transcriptional coactivators that have been implicated in driving metastasis and progression in many cancers, mainly through their transcriptional regulation of downstream targets. Although YAP and TAZ have shown redundancy in many contexts, it is still...
The failure of once promising target-specific therapeutic strategies often arises from redundancies in gene expression pathways. Even with new melanoma treatments, many patients are not responsive or develop resistance, leading to disease progression in terms of growth and metastasis. We previously discovered that the transcription factors ETS1 and...
Transcriptional regulation is a tightly regulated, vital process. The transcription factor (TF) CREB1 controls ~25% of the mammalian transcriptome by binding the CRE sequence (TGACGTCA). DNA lesions within CRE modulate CREB1 binding negatively and positively. Because appropriate DNA lesions also interact with base excision repair (BER) proteins, we...
CREB controls ∼25% of the mammalian transcriptome. Small changes in binding to its consensus (CRE) sequence are likely to be amplified many fold in initiating transcription. Here we show that DNA lesions repaired by the base excision repair (BER) pathway modulate CREB binding to CRE. We generated Kd values by electrophoretic mobility shift assays u...
I'm currently trying to run MNase digestion assays using zebrafish embryos. I have all my protocols down and working well, only problem is I don't seem to be able to quantify the nuclei/DNA so that I can normalise across tubes. Most papers/protocols seem to state that they quantified using A260. I've tried taking sub-samples of my resuspended nuclei and quantifying at A260 on a nanodrop, but don't get any consistency. There is also a lot of interference from other components. I also tried lysing nuclei in the sub-sample to make the DNA more accessible, but still problems. Any help or advice would be much appreciated.