Journal: Cell chemical biology

Article Title: Discovery of covalent CDK14 inhibitors with pan-TAIRE family specificity

doi: 10.1016/j.chembiol.2019.02.015

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: RB , Signal Chem , Cat # R05-55G-50.

Techniques: Virus, Recombinant, Kinase Assay, Sequencing, Software

Journal: Nature cell biology

Article Title: An androgen receptor switch underlies lineage infidelity in treatment-resistant prostate cancer

doi: 10.1038/s41556-021-00743-5

Figure Lengend Snippet: a, Heat map depicting normalized z-score for genes in the pEZH2-T350 signature in NPp53 GEMMs (Gene Expression Omnibus (GEO) accession: GSE92721). ABI, abiraterone acetate; NR, exceptional non-responder. b, pEZH2-T350 signature score in patient tumours from Aggarwal (ref. 5) (P = 0.0019) and Beltran (ref. 4) (P = 0.012). Violin plots show mean and interquartile range, with significance assessed using a two-tailed unpaired t-test. AdPC, prostate adenocarcinoma; tNEPC, treatment-induced neuroendocrine prostate cancer. c, Correlation between pEZH2-T350 and ASC scores in NEPC tumours from ref. 4. Each dot represents a patient tumour showing RB1 loss-of-function score. d, Immunohistochemical staining in patient-derived NEPC organoids. Scale bar, 100 μm. e, Immunohistochemical staining in PB-Cre4:Ptenf/f and PB-Cre4:Ptenf/f;Rb1f/f GEMMs. Scale bars: 200 μm (prostate), 1 mm (liver), 20 μm (insets). f, pEZH2-T350 score in PTEN-null GEMMs with RB1 deletion alone (P = 0.022) or concomitant with TP53 loss (P = 0074; GEO accession: GSE90891). Dots represent individual tumours. Box plot shows mean and interquartile range, with significance assessed using a two-tailed unpaired t-test. g, Volcano plot of RNA-seq from PB-Cre4:Ptenf/f and PB-Cre4:Ptenf/f;Rb1f/f GEMMs (GEO accession: GSE90891). Each dot represents a gene, with those in the ‘Reactome neuronal system’ (neuronal) and ‘Wong embryonic stem cell core’ (plasticity) MSigDB pathways highlighted. h, Immunoblot in 16DCRPC cells with stable RB1 knockdown treated with CDK1 inhibitor (5 μM RO-3306, 6 h). shNS, non-silencing control. i, Heat map of gene expression (by rtPCR) in CRPCcrEZH2 cells with shRB1 expressing EZH2WT, EZH2T350A or EZH2T350D mutant constructs following treatment with DMSO or ENZ (10 μM) for 7 d. Expression is reported relative to parental cells (n = 3). j, Left: quantification of spheroids in CRPCcrEZH2 cells with shRB1 expressing EZH2T350A or EZH2T350D treated with DMSO or ENZ (10 μM). Data reported as mean ± s.d., with significance evaluated at the end point (P = 0.003, two-tailed unpaired t-test; n = 3). Right: images at 5 d after ENZ treatment. Scale bar, 100 μm. k, Left: neurite length measured using IncuCyte in CRPCcrEZH2 cells with shRB1 expressing EZH2T350A or EZH2T350D and treated with DMSO or ENZ (10 μM). Right: phase-contrast images at 96 h after ENZ treatment, with neurite-like extensions highlighted by NeuroTrack. Data reported as mean ± s.d. with significance evaluated at end-point (P = 0.0003, two-tailed unpaired t-test; n = 3). Scale bar, 100 μm.

Article Snippet: The shRB1 (#25641), pcDNA3-EZH2 S21A (#42663), pcDNA3-EZH2 S21D (#42664) and control (#8453) plasmids were available from Addgene.

Techniques: Expressing, Two Tailed Test, Immunohistochemical staining, Staining, Derivative Assay, RNA Sequencing Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction, Mutagenesis, Construct