Journal: bioRxiv

Article Title: Effects of breast fibroepithelial tumor associated retinoic acid receptor alpha ligand binding domain mutations on receptor function and retinoid signaling

doi: 10.1101/2022.12.01.518133

Figure Lengend Snippet: Mutant RARα possess altered binding abilities to agonists and co-regulators. Mammalian two-hybrid assay of interactions of the wild type or mutant RARα LBD with (A) fragment of coactivator SRC2, (B) MED1, a component of the transcription pre-initiation complex and (C) fragment of corepressor NCOR, treated with increasing concentrations of RA. Left panel: Partially activatable mutants, Right panel: Non-activatable mutants. N=3, Error bar = SD. (D) Binding affinities of WT or mutant RARα LBD with fragment of coactivator SRC1 (SRC1-NR2) in the presence of RA and Am80. N=3, error bar = SE. **p<0.01.

Article Snippet: Compounds tested include all-trans retinoic acid (known as RA throughout the manuscript, Sigma R2625), Am80 (aka Tamibarotene, Tocris 3507), AM580 (Tocris 0760) and Ch55 (Tocris 2020).

Techniques: Mutagenesis, Binding Assay, Two Hybrid Assay

Journal: bioRxiv

Article Title: Effects of breast fibroepithelial tumor associated retinoic acid receptor alpha ligand binding domain mutations on receptor function and retinoid signaling

doi: 10.1101/2022.12.01.518133

Figure Lengend Snippet: Sensitivity of mutant RARα to retinoid treatment. (A) Overexpression of Flag-tagged RARα WT and mutants in PT024 cells. (B) Cell viabilities of PT024 WT or mutant RARα expressing cells after treatment with increasing concentration of RA or Am80. N=3, error bar = SD. ***p<0.001, ****p<0.0001. (C) Cell cycle distribution of PT024 WT or mutant RARα expressing cells when treated with either vehicle or 100nM Am80. N=2, error bar = SD. *p<0.05, ****p < 0.0001. (D) GSEA analysis shows downregulation of retinoic acid signaling gene targets in mutant RARα-expressing cell lines. (E) Heatmap of genes associated with reactome retinoic acid signaling pathway (F) Quantitative PCR of retinoic acid signaling associated genes. N=3, error bar = SD. *p<0.05 **p<0.01 ***p<0.001 ****p < 0.0001.

Article Snippet: Compounds tested include all-trans retinoic acid (known as RA throughout the manuscript, Sigma R2625), Am80 (aka Tamibarotene, Tocris 3507), AM580 (Tocris 0760) and Ch55 (Tocris 2020).

Techniques: Mutagenesis, Over Expression, Expressing, Concentration Assay, Real-time Polymerase Chain Reaction

Journal: F1000Research

Article Title: An assay to measure poly(ADP ribose) glycohydrolase (PARG) activity in cells

doi: 10.12688/f1000research.8463.2

Figure Lengend Snippet: Using a high content imaging system the Hoechst stained nuclei (left-hand panels) are used to create a nuclear mask (green circle in Analysis). Anti-PAR antibody (FITC) detects the increase in PAR chains (centre panels) that is then quantified using the nuclear mask from the Hoechst signal (right-hand panels). Fluorescence intensity is shown as red dots within the nuclear mask.

Article Snippet: Cells were then permeabilized using PBS/Triton 0.1% for 20 min, and washed once in PBS before adding anti-PAR antibody (10H (#AM80), Merck Millipore) at 1:4000 in antibody blocking buffer (ADB; 5% Fetal bovine serum, 0.1% Tween20 in PBS) and incubated overnight at 4°C.

Techniques: Imaging, Staining, Fluorescence

Journal: PLoS ONE

Article Title: Retinoic Acid Upregulates Ret and Induces Chain Migration and Population Expansion in Vagal Neural Crest Cells to Colonise the Embryonic Gut

doi: 10.1371/journal.pone.0064077

Figure Lengend Snippet: Abbreviations as in , plus RA = retinoic acid; AM80 = synthetic retinoid which is not able to be processed by CYP26A1.

Article Snippet: For different conditions (detailed in results) soluble reagents were added to the NCC donor media either before placement with the gut or during culture, at the following concentrations: 10 µM all-trans-RA (Sigma, USA), 10 µM Am80 synthetic retinoid (Wako Chemicals USA, Inc).

Techniques: