br Experiment hydrogen peroxide exposure with SH

Experiment 4: hydrogen peroxide exposure with SH-SY5Y neurons SH-SY5Y neuronal wnt signaling pathway were cultured without experimental perturbation for a minimum of 5 passages. Cells were then changed to media containing either 200nM or 0nM H2O2 for 2h at least 24h after the previous media change. Cells were then fixed and stained as described above. See Fig. 5.
Sources of funding Sources of funding for this study were provided by VA Merit, Denver Research Institute, NIH-5T32HL007171 (ACK), NIH-5P01HL014985 (ACK), CCTSI-UL1RR025780 (JEBR), the Center for Women’s Health Research (JEBR), and the Department of Bioengineering (PMM). Flow Cytometry is supported through a National Cancer Institute Cancer Center Support Grant (P30CA046934).
Disclosures
Acknowledgments
Specifications table
Value of the data
Data High-content screening allows quantification of data obtained by fluorescence imaging in a multi-well format. Using this technology, we were able to statistically differentiate substrate levels (p<0.0001) between normal (GLB1+/+) and enzyme deficient (GLB1−/−) human fibroblasts. Reduction of substrate levels can be detected when GM1-gangliosidosis fibroblast (GLB1−/−) are treated with a corrective recombinant protein.
Experimental design, materials and methods
Acknowledgments This research was supported by funding to BioStrategies LC from an NIH/NINDS Phase I SBIR grant (1R43NS084565-01). The BD pathway was purchased through a NSF MRI grant (DBI-0960089) to C.L.C, Arkansas State University.
Value of the data
Data The protein content and DLS count-rate events of EVs isolated from MDR and drug-sensitive cells have been shown (Table 1). In Table 2 a list of proteins identified by mass spectrometry, in EVs isolated from CML cells (K562 – drug-sensitive cells and K562Dox – MDR cells) have been mentioned. Moreover, Fig. 1 shows Gene Ontology analysis performed in the protein list obtained by LC/MS/MS of EVs isolated from K562 and K562Dox cells, based on the biological processes, molecular functions and pathways.
Experimental design, materials and methods EVs were isolated from two pairs of isogenic cell lines (MDR and the drug-sensitive counterpart) from two different cancer models, non-small cell lung cancer-NSCLC (H460 – drug-sensitive cells and RH460 – MDR cells)[2,3] and chronic myeloid leukaemia-CML (K562 – drug-sensitive cells and K562Dox – MDR cells) [4,5]. Cells were used to isolate EVs, for protein quantification (Table 1), Dynamic light scattering (Table 1) and proteomic experiments (Table 2 and Fig. 1).
Acknowledgements IPATIMUP integrates the i3S Research Unit, which is partially supported by FCT, the Portuguese Foundation for Science and Technology. This work is funded by FEDER funds through the Operational Programme for Competitiveness Factors-COMPETE and National Funds through the FCT-Foundation for Science and Technology, under the projects “PEst-C/SAU/LA0003/2013” NORTE-07-0162-FEDER-00018 – Contributos para o reforço da capacidade do IPATIMUP enquanto actor do sistema regional de inovação and NORTE-07-0162-FEDER-000067-Reforço e consolidação da capacidade infraestrutural do IPATIMUP para o sistema regional de inovação, both supported by Programa Operacional Regional do Norte (ON.2 – O Novo Norte), through FEDER funds under the Quadro de Referência Estratégico Nacional (QREN). The proteomic work was also made possible through funding provided in part from awards from Science Foundation Ireland, Grant code 08/SRC/B1410 and the Irish Cancer Society, Grant code CCRC13GAL. The authors thank the Portuguese Foundation for Science and Technology (FCT) for the PhD grants of VLR and DS (SFRH/BD/87646/2012 and SFRH/BD/98054/2013, respectively) and for the post-doc grant of RTL (SFRH/BPD/68787/2010).
Data Data presented here describes the additional chemical analysis of the “krokodil” samples obtained using the street-like synthesis. Physical and organoleptic characters, UV/Vis and 1H NMR spectra were described on a “krokodil” sample freshly prepared (crude “krokodil”). Organic extract of “krokodil” (extracted “krokodil”) was obtained after alkalization of the crude product and extraction using ethyl acetate. This organic extracts were analyzed by TLC, FTIR and 1H NMR techniques.