5 research outputs found
Analysis of <i>Plagl1</i> region in <i>Dnmt3l ā/+.</i>
<p>(A) The methylation status of the <i>Plagl1</i> promoter regions in wild type +/+ and <i>Dnmt3l</i> ā/+ embryos examined by bisulphite PCR. Each circle represents a single CpG dinucleotide on a DNA strand, a methylated cytosine (ā¢) or an unmethylated cytosine (ā). (B) RT-PCRs on cDNA generated with (+) and without (ā) reverse transcriptase show an increase in the expression of the imprinted transcripts in <i>Dnmt3l</i>ā/+ embryos as a result of reactivation of the maternal allele. (C) The histone modification signature of the <i>Plagl1</i>-DMR in wild type BĆC embryos, and after targeted deletion of the <i>Dnmt3l</i> gene. DNA extracted from antibody bound (B) and unbound (U) chromatin fractions were subject to either qPCR or PCR and SSCP analysis with primers that can discriminate parental alleles.</p
Cellular localization and RNA stability of the ncRNAs.
<p>(A) Distribution of the various transcripts in the nuclear (dark grey) and cytoplasmic (black) fractions, compared to total RNA (light grey). <i>U937 snoRNA</i> and <i>Airn</i> are nuclear-retained controls, whereas <i>Igf2</i> is cytoplasm-exported control. (B) Abundance of the various transcripts after exposure to Actinomycin D to determine RNA stability. The relative expression values of the control untreated samples are set to 1 (light grey bars) for each transcript. <i>C-Myc</i> and <i>Airn</i> are control transcripts for with short half-life; <i>Gapdh</i> and <i>Igf2r</i> are long half-life controls.</p
Schematic overview of the mouse chromosome 10 imprinted domain.
<p>(A) Map of the <i>Plagl1</i> locus, showing the location of the various imprinted transcripts and CpG islands (paternally expressed transcripts are in blue; biallelically expressed transcripts are in grey). Arrows represent direction of transcription. (B) The allelic expression of the various transcripts in embryonic tissues in reciprocal mouse crosses (for clarity only (BĆC) F1 tissues are shown).</p
Structural Basis for Highly Selective Class II Alpha Phosphoinositide-3-Kinase Inhibition
Class
II phosphoinositide-3-kinases (PI3Ks) play central roles
in cell signaling, division, migration, and survival. Despite evidence
that all PI3K class II isoforms serve unique cellular functions, the
lack of isoform-selective inhibitors severely hampers the systematic
investigation of their potential relevance as pharmacological targets.
Here, we report the structural evaluation and molecular determinants
for selective PI3K-C2Ī± inhibition by a structureāactivity
relationship study based on a pteridinone scaffold, leading to the
discovery of selective PI3K-C2Ī± inhibitors called PITCOINs.
Cocrystal structures and docking experiments supported the rationalization
of the structural determinants essential for inhibitor activity and
high selectivity. Profiling of PITCOINs in a panel of more than 118
diverse kinases showed no off-target kinase inhibition. Notably, by
addressing a selectivity pocket, PITCOIN4 showed nanomolar
inhibition of PI3K-C2Ī± and >100-fold selectivity in a general
kinase panel. Our study paves the way for the development of novel
therapies for diseases related to PI3K-C2Ī± function
Structural Basis for Highly Selective Class II Alpha Phosphoinositide-3-Kinase Inhibition
Class
II phosphoinositide-3-kinases (PI3Ks) play central roles
in cell signaling, division, migration, and survival. Despite evidence
that all PI3K class II isoforms serve unique cellular functions, the
lack of isoform-selective inhibitors severely hampers the systematic
investigation of their potential relevance as pharmacological targets.
Here, we report the structural evaluation and molecular determinants
for selective PI3K-C2Ī± inhibition by a structureāactivity
relationship study based on a pteridinone scaffold, leading to the
discovery of selective PI3K-C2Ī± inhibitors called PITCOINs.
Cocrystal structures and docking experiments supported the rationalization
of the structural determinants essential for inhibitor activity and
high selectivity. Profiling of PITCOINs in a panel of more than 118
diverse kinases showed no off-target kinase inhibition. Notably, by
addressing a selectivity pocket, PITCOIN4 showed nanomolar
inhibition of PI3K-C2Ī± and >100-fold selectivity in a general
kinase panel. Our study paves the way for the development of novel
therapies for diseases related to PI3K-C2Ī± function