Journal: Nature

Article Title: Discovery and characterization of small molecules that target the Ral GTPase

doi: 10.1038/nature13713

Figure Lengend Snippet: Ras and Ral knockdown in human cancer cell lines a–b , Effect of K-Ras knockdown on anchorage-independent growth of four human lung cancer cell lines. a , Immunoblot showing siRNA knockdown of K-Ras in H2122, H358, H460, and Calu6 cell lines 48h after siRNA transfection. b , All four lines were sensitive to K-Ras knockdown using the soft agar colony formation assay. Data represents the mean ± SD of triplicate samples. * Statistically different from respective controls as determined by the Student’s t-test (p

Article Snippet: Human bladder cancer cell line J82 and lung cancer cell lines H2122, H358, H460, and Calu6 were obtained from ATCC.

Techniques: Transfection, Soft Agar Assay

Journal: Nature

Article Title: Discovery and characterization of small molecules that target the Ral GTPase

doi: 10.1038/nature13713

Figure Lengend Snippet: Ral inhibitors on human cancer cell lines in vitro a–c , Cellular uptake of Ral inhibitors in vitro . H2122 human lung cancer cells were treated with RBC5, RBC8 and BQU57 (10 μM). Cells were collected at various time points (1, 5, 15, 30 and 60 min), and drug concentrations in cells determined using LC/MS-MS methods. Data represents the mean ± SD of triplicate samples. d , Effect of RBC5 treatment on the anchorage-independent growth of H2122 and H358 human lung cancer cell lines. Cells were seeded in soft agar containing various concentrations of drug; colonies formed in soft agar were counted after 2–4 weeks. Data represents the mean ± SD of triplicate samples. e , Inhibition of Ral activity in H2122 and H358 cells by RBC5, RBC8 and BQU57. Cells were grown under anchorage-independent conditions and treated with 10 μM compounds for 3 hrs. Ral activity in cell lysates were then determined using the pull down assay with RalBP1 agarose beads. Total lysates (20 μg protein) and RalBP1 pull-downs (from 400 μg protein) were analyzed by immune-blotting using antibodies specific for RalA and RalB. Data represent three independent experiments.

Article Snippet: Human bladder cancer cell line J82 and lung cancer cell lines H2122, H358, H460, and Calu6 were obtained from ATCC.

Techniques: In Vitro, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Inhibition, Activity Assay, Pull Down Assay

Journal: Nature

Article Title: Discovery and characterization of small molecules that target the Ral GTPase

doi: 10.1038/nature13713

Figure Lengend Snippet: Effect of Ral inhibitors on human xenograft models of lung cancer a , Summary of pharmacokinetic parameters of RBC8 and BQU57 in nu/nu mice. Animal PK parameters were measured based on plasma levels after 50 mg/kg single intraperitoneal dose. Abbreviations used: C o , extrapolated initial concentration; T 1/2 , half-life; AUC 0–5hr , area under the curve, zero to five hours. b–c , Tissue distribution of RBC8 ( b ), BQU57 ( c ) in nude mice 3h after a single i.p. dose of 50 mg/Kg of drug. Data represent the mean ± SD of 3 mice. d , 50 mg/kg/day RBC8 initiated 24h after inoculation inhibited xenograft tumor growth of human lung cancer cell line H2122. Typical tumor appearance at 21 days are shown. e , Effect of RBC8 on H358 xenograft models. RBC8 treatment (50 mg/kg/day) initiated 24 h after inoculation inhibited xenograft tumor growth of human lung cancer cell line H358. Data represents the mean ± SEM of 6 mice. Tumor volume in the treatment group was statistically different from controls as determined by the Student’s t-test (*p

Article Snippet: Human bladder cancer cell line J82 and lung cancer cell lines H2122, H358, H460, and Calu6 were obtained from ATCC.

Techniques: Mouse Assay, Concentration Assay

Journal: BMC Cancer

Article Title: p300 promotes proliferation, migration, and invasion via inducing epithelial-mesenchymal transition in non-small cell lung cancer cells

doi: 10.1186/s12885-018-4559-3

Figure Lengend Snippet: a Relative p300 expression of nine aggressive non-small cell lung cancer cell lines were examined with Western Blot analysis; b NCI-H1975 and NCI-H1993 cells were transfected with shp300 and shNC, western blot was used to determine interference efficiency; c P300-pcDNA3.1-EGFP and scrambled plasmid were transfected into NCI-H460, western blot was used to determine transfection efficiency

Article Snippet: The human NSCLC cell lines NCI-H292 (ATCC CRL-1848), NCI-H460 (ATCC HTB-177), PC-9 (RRID:CVCL_B260), A549 (ATCC CCL-185), NCI-H1650 (ATCC CRL-5883), NCI-H1993 (ATCC CRL-5909), NCI-H1975 (ATCC CRL-5908), HCC827 (ATCC CRL-2868), and NCI-H1299 (ATCC CRL-5803) were obtained from the State Key Laboratory (SKL) of Oncology in South China.

Techniques: Expressing, Western Blot, Transfection, Plasmid Preparation