The mesothelin (MSLN) gene offers a promising subject, being expressed in a restricted pattern normally, yet highly overexpressed

in almost one-third of human malignancies and a target of cancer immunotherapeutic trials. CanScript, a cis promoter element, appears to control MSLN cancer-specific expression; its related genomic sequences may up-regulate other cancer markers. CanScript is a 20-nt bipartite element consisting of an SP1-like motif and a consensus MCAT sequence. The latter recruits TEAD (TEA domain) family members, which are universally expressed. Exploration of the active CanScript element, especially the proteins binding to the SP1-like motif, thus could reveal cancer-specific features having diagnostic or therapeutic interest. The efficient identification of sequence-specific DNA-binding proteins at a given locus, selleck chemical however, has lagged in biomarker explorations. We used two orthogonal proteomics approaches-unbiased SILAC (stable isotope labeling by amino acids in cell culture)/DNA affinity-capture/mass spectrometry survey (SD-MS) and a large transcription factor protein microarray (TFM)-and functional validation to explore systematically the CanScript interactome. SD-MS produced nine

candidates, and TFM, 18. The screens agreed in confirming binding by TEAD proteins and by newly identified NAB1 and NFATc. Among other identified Baf-A1 candidates, we found functional roles for ZNF24, NAB1 and RFX1 in GDC-0068 nmr MSLN expression by cancer cells. Combined interactome screens yield an efficient, reproducible, sensitive, and unbiased approach to identify sequence specific DNA-binding proteins and other participants in disease-specific DNA elements.”
“Three new clerodane diterpene glycosides, tinospinosides A (1), B (2), and C (3) were isolated from the roots of Tinospora sagittata (Oliv.) Gagnep. Their structures were determined to be (2S, 4aR, 6aR,

9R, 10aS, 10bS)-2-(3-furanyl)-9-(beta-D-glucopyranosyloxy)-1,4,4a, 5,6,6a, 9,10,10a, 10b-decahydro-6a, 10b-dimethyl-4oxo- 2H-naphtho[2,1-c] pyran-7-carboxylic acid methyl ester (1), (2S, 4aS, 6aR, 9R, 10aR, 10bS)-2-(3-furanyl)-9-(beta-D-glucopyranosyloxy)- 1,4,4a, 5,6,6a, 9,10,10a, 10b-decahydro-4a-hydroxyl-6a, 10b-dimethyl- 4-oxo-2H-naphtho[2,1-c] pyran-7-carboxylic acid methyl ester (2) and (2S, 4aR, 6aR, 9R, 10aR, 10bS)-2-(3-furanyl)-9-(beta-D- glucopyranosyloxy)-1,4,4a, 5,6,6a, 9,10,10a, 10b-decahydro-4a-hydroxyl- 6a, 10b-dimethyl-4-oxo-2H-naphtho[2,1-c] pyran-7carboxylic acid methyl ester (3), by various spectroscopic analyses, chemical reactions, and computer-assisted calculations. The inhibitory activities of NO production by these compounds and their chemical derivatives in lipopolysaccharide and TNF gamma-activated macrophage-like cell line J774.1 were tested. Tinospin A, 12-epi-tinospin A, tinospinoside B, and tinospinoside C showed inhibitory activities of NO production with the IC(50) values of 162, 182, 290, and 218 mu M, respectively.