Journal: Journal of Neuroscience

Article Title: Specific Acetylation of p53 by HDAC Inhibition Prevents DNA Damage-Induced Apoptosis in Neurons

doi: 10.1523/jneurosci.5214-12.2013

Figure Lengend Snippet: Figure 8. Acetylation of lysines 381 and 382 promotes the pro-apoptotic functions of p53 in proliferating tumor cell lines. A, HCT116, DLD1, and HT22 cells were transfected with an equal amount of plasmid DNA encoding wild-type human p53 (WT), DNA-bindingmutantp53(R175H),orthep53acetylationmutantsK373QandK381/382Q.Immunoblotanalysiswasperformedon total cell lysates 24 h after electroporation using antibodies specific for human p53. Plasmid DNA encoding eGFP was used as a transfection control. B, Luciferase reporter assay of plasmid DNA encoding a fragment of the human PUMA proximal promoter (PUMAFrag1-Luc)inhumancolorectalcarcinomacelllinesHCT116andDLD-1andinmousehippocampalcelllineHT22.Promoter reporterplasmidwascotransfectedwithDNA-bindingmutantp53(R175H),wild-typep53(WT),K373Q,orK381/382Qacetylation mutants of p53. Error bars indicate SD of mean from three independent experiments. ***p 0.001, significant difference compared with R175H; ###p 0.001, ##p 0.01, significant difference compared with WT (one-way ANOVA followed by Bonferroni’stest).C,CellviabilitymeasuredbyMTTassayofHCT116,DLD-1,andHT22cultures24haftertransfectionwithR175H, WT, K373Q, or K381/382Q acetylation mutants of p53. Error bars indicate SD of mean from three independent experiments. ***p0.001,**p0.01,*p0.05,significantdifferencecomparedwithR175H; ##p0.01,significantdifferencecompared withWT(one-wayANOVAfollowedbyBonferroni’stest).D,HCT116celldeathassessedbyethidiumhomodimerstaining(EthD-1; red) 24 h after transfection with R175H, WT, K373Q, or K381/382Q acetylation mutants of p53.

Article Snippet: DLD-1 and HCT116 human cell lines were purchased from American Type Culture Collection.

Techniques: Transfection, Plasmid Preparation, Electroporation, Control, Luciferase, Reporter Assay

Journal: Journal of Neuroscience

Article Title: Specific Acetylation of p53 by HDAC Inhibition Prevents DNA Damage-Induced Apoptosis in Neurons

doi: 10.1523/jneurosci.5214-12.2013

Figure Lengend Snippet: Figure 9. HDAC inhibition does not prevent PUMA induction and DNA damage-induced death in HCT116 cells.A, Immunoblot analysisofpan-p53,phosphorylatedp53(pS15-p53),andacetylatedp53(AcK381-p53andAcK382-p53)inextractsfromHCT116 cells treated with CPT (20 M) with or without TSA (1.3 M) for 8 h. B, Quantitative RT-PCR of PUMA expression in HCT116 cells treatedwithCPT(20M)withorwithoutTSA(1.3M)for8h.ErrorbarsindicateSDofmeanfromthreeindependentexperiments. ***p 0.001, significant difference compared with untreated control; ##p 0.01, significant difference compared with CPT (one-wayANOVAfollowedbyBonferroni’stest).C,HCT116cellviabilitymeasuredbyMTTassayaftertreatmentwithCPT(20M) with or without TSA (1.3 M). Error bars indicate SD of mean from three independent experiments. ***p 0.001, **p 0.01, significant difference compared with untreated control; #p 0.05, significant difference compared with CPT (one-way ANOVA followedbyBonferroni’stest).D,HCT116celldeathassessedbyethidiumhomodimerstaining(EthD-1;red)aftertreatmentwith CPT (20 M) with or without TSA (1.3 M).

Article Snippet: DLD-1 and HCT116 human cell lines were purchased from American Type Culture Collection.

Techniques: Inhibition, Western Blot, Quantitative RT-PCR, Expressing, Control

Journal: British Journal of Cancer

Article Title: Loss of Coxsackie and adenovirus receptor downregulates α -catenin expression

doi: 10.1038/sj.bjc.6605331

Figure Lengend Snippet: Impact of CAR and α -catenin on cellular morphology. Cells after CAR knockdown appear round and smaller compared with controls (DLD1) or display less intercellular attachment sites (IEC-6). In matrigel, cells after CAR knockdown form amorphous clusters in contrast to the organised-appearing formations of matching controls. Ectopic ‘re’-expression of α -catenin partially reverses the effects seen after CAR knockdown when grown as a monolayer, and particularly results in organised cell formations similar to those of vector controls when cultured in matrigel. Images show representative results for DLD1 ( A ) and IEC-6 ( B ).

Article Snippet: The human colon cancer cell line DLD1 and the rat small intestine cell line IEC-6 were obtained from the American Type Culture Collection (Rockville, MD, USA) and from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (Braunschweig, Germany), respectively, and were cultured in recommended growth media.

Techniques: Knockdown, Expressing, Plasmid Preparation, Cell Culture

Journal: British Journal of Cancer

Article Title: Loss of Coxsackie and adenovirus receptor downregulates α -catenin expression

doi: 10.1038/sj.bjc.6605331

Figure Lengend Snippet: Functional impact of CAR downregulation and α -catenin ‘re’-expression in DLD1 and IEC-6 cell lines. Proliferation ( A ), migration ( B ), and invasion ( C ) in DLD1 and IEC-6 were determined after CAR knockdown and ‘re’-expression of α -catenin in comparison with controls.

Article Snippet: The human colon cancer cell line DLD1 and the rat small intestine cell line IEC-6 were obtained from the American Type Culture Collection (Rockville, MD, USA) and from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (Braunschweig, Germany), respectively, and were cultured in recommended growth media.

Techniques: Functional Assay, Expressing, Migration, Knockdown, Comparison