Project description:LC-MS and GC-MS raw data for 13C6-glucose tracing in PDA cells expressing sgROSA or sgRNA targeting methionine sulfoxide reductase A
Project description:Pancreatic ductal adenocarcinoma (PDA) is characterized by abundant desmoplasia and poor tissue perfusion. These features are proposed to limit access of therapies to neoplastic cells and blunt treatment efficacy. Indeed, several agents that target the PDA microenvironment promote chemotherapy delivery and improve anti-neoplastic responses in murine models of PDA. Here, we employed the FG-3019 monoclonal antibody directed against the pleiotropic matricellular signaling molecule connective tissue growth factor (CTGF/CCN2). FG-3019 treatment increased PDA cell killing and led to a dramatic tumor response without altering gemcitabine delivery. Microarray expression profiling revealed the down-regulation by FG-3019 of several anti-apoptotic transcripts, including the master regulator Xiap, down-regulation of which has been shown to sensitize PDA to gemcitabine. Decreases in XIAP protein by FG-3019 in the presence and absence of gemcitabine were confirmed by immunoblot, while increases in XIAP protein were seen in PDA cell lines treated with recombinant CTGF. Therefore, alterations in survival cues following targeting of tumor microenvironmental factors may play an important role in treatment responses in animal models and, by extension, PDA patients. Total RNA was isolated from KPC mouse PDA tumors 9 days after initiation of treatment with IgG (n=7 biological replicates), FG-3019 (n=5), IgG + gemcitabine (n=6), or FG-3019 + gemcitabine (n=6) and hybridized to Affymetrix 430A 2.0 microarrays. CEL files were processed by GC-RMA and rescaled using median per-gene normalization in GeneSpring GX 7.3.1.
Project description:22 week old female Tfamfl/fl (aka WT), BEST1Cre;Tfamfl/fl, BEST1Cre;Tfamfl/fl;AN/+ mice were fasted for 6 hrs (8am-2pm), then were retro-orbital injected with 13C6-glucose (Cambridge Isotopes Laboratories): 250mg/kg in 100-135ul saline and 0.2 um sterile filtered. Tissues were processed 30min after injection. A cryolysis method was used to extract RPE metabolites. NR metabolites were extracted as usual.
Project description:Human cancer cell lines indicated in the file names, with stably overexpressed Cas9 nuclease, were transfected with (C) control sgRNA or (P) TP53-specific sgRNA, (K) KRAS-specific sgRNA or (M) CMYC-specific sgRNA. Each experiment was done by transfection of sgRNA pair - control (C) or causing a NHEJ-mediated knock-out of the target genes (P, K or M). Samples for proteomics were collected 48h post sgRNA transfection without selection, in three biological replicates each (indicated with numbers 1-3). In cell lines with three activated oncogenes, three separate oncogene-targeting transfections were carried out.
Project description:This project provides tandem mass spectrometry datasets of Bacillus cereus ATCC 14579 wild-type strain without its pBClin15 plasmid devoided of both the methionine sulfoxide reductase AB (msrAB)and methionine sulfoxide reductase A (msrA) genes. The double mutant strain was grown under fermentative anaerobic condition and harvested at three growth stages.
Project description:This project provides tandem mass spectrometry datasets of Bacillus cereus ATCC 14579 wild-type strain without its pBClin15 plasmid devoided of both the methionine sulfoxide reductase AB (msrAB)and the methionine sulfoxide reductase A (msrA) genes. The double mutant strain was grown under fermentative anaerobic condition and harvested at three growth stages.
Project description:The HS766T PDA cell line was used to experimentally assess cross-lineage plasticiyty between different states of Pancreatic Malignant Cells