• 2019-10
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  • br Analysis of tumor growth br Luciferase


    5.9. Analysis of tumor growth
    Luciferase-positive GBM tumors were imaged using an In Vivo Imaging System (IVIS) Spectrum (Caliper Life Sciences) and analyzed using Living Image® software (PerkinElmer). Animals were injected intraperitoneally with 4.5 mg D-luciferin potassium salt (GoldBio) in 1 × PBS 10 min before imaging. Mice (n = 5 at experiment start date)
    5.10. Analysis of PF 6700841 tissue and tumor size
    Four weeks after tumor inoculation, animals were anesthetized using ketamine/xylazine and perfused with 10% formalin. The skulls were removed and brains stored in 10% formalin overnight. Brains were immersed in 30% sucrose for two days and then in Optimal Cutting Temperature (OCT, Fisher Scientific) overnight. The brains in OCT were frozen and sectioned with a Leica CM1905 cryostat (14-μm sections). Six sections from each animal (n = 3 mice per group) were stained with hematoxylin and eosin (H&E), and the slides were imaged with a Zeiss Axio observer A1 microscope with a Zeiss Axiocam MRm camera PF 6700841 at 2.5 × magnification. The tumor area in each H&E-stained section was measured via computer-assisted morphometric quantifica-tion using AxioVision 4.8 software.
    5.11. Statistical analysis
    Unless otherwise indicated, the data are presented as the mean ± standard error of the mean. In vitro siRNA nanoparticle uptake and transfection efficacy in GBM versus NPC cells and differences in tumor burden between groups were analyzed using a two-tailed t-test. One-way ANOVA with Dunnett's post-tests were used to evaluate multiple comparisons to a control group and a Bonferroni correction was used when comparing a greater number of groups. p-values < 0.05 were considered statistically significant in all cases.
    Conflicts of interest
    The authors declare no conflict of interest.
    The authors would like to thank Montserrat Lara-Velazquez, Paola Suarez-Meade, Carla Vazquez-Ramos, and Emily Lavell, for assistance with animal surgery and histological processing, Rachel Sarabia-Estrada for assistance with animal surgery and the use of surgical equipment, and Barbara Kim for assistance with transfections and Western blotting. The authors thank the NIH (R01EB016721, R01CA195503, and R01CA228133) for support of this work. AQH is also supported by the Mayo Clinician Investigator Award, Mayo Professorship, and the State of Florida. HGC is also supported by the NCI (5R21CA199295). JJG thanks the Bloomberg ∼ Kimmel Institute for Cancer Immunotherapy for support. KLK thanks the National Cancer Institute (NIH F31CA196163), and the ARCS Foundation for fellowship support. JK thanks Samsung for scholarship support.
    Data Availability
    The raw/processed data required to reproduce these findings cannot be shared at this time as the data also forms part of an ongoing study.
    Appendix A. Supplementary data
    [4] K.L. Chaichana, I. Jusue-Torres, R. Navarro-Ramirez, S.M. Raza, M. Pascual-Gallego, A. Ibrahim, M. Hernandez-Hermann, L. Gomez, X. Ye, J.D. Weingart, Establishing percent resection and residual volume thresholds affecting survival and recurrence for patients with newly diagnosed intracranial glioblastoma, Neuro Oncol. 16 (1) (2013) 113–122.
    S. Dangelmajer, H. Guerrero-Cazares, A. Quinones-Hinojosa, S.A. Goldman, PAR1 inhibition suppresses the self-renewal and growth of A2B5-defined glioma pro-genitor cells and their derived gliomas in vivo, Oncogene 35 (29) (2016)
    [15] K.L. Kozielski, S.Y. Tzeng, B.A.H. De Mendoza, J.J. Green, Bioreducible cationic polymer-based nanoparticles for efficient and environmentally triggered cyto-plasmic siRNA delivery to primary human brain cancer cells, ACS Nano 8 (4) (2014) 3232–3241.
    [21] P. Schiapparelli, H. Guerrero-Cazares, R. Magana-Maldonado, S.M. Hamilla,
    S. Ganaha, E. Goulin Lippi Fernandes, C.H. Huang, H. Aranda-Espinoza,
    P. Devreotes, A. Quinones-Hinojosa, NKCC1 regulates migration ability of glio-blastoma cells by modulation of actin dynamics and interacting with cofilin, EBioMedicine 21 (2017) 94–103.
    [26] A.Y. Galvez-Contreras, A. Quiñones-Hinojosa, O. Gonzalez-Perez, The role of EGFR and ErbB family related proteins in the oligodendrocyte specification in germinal niches of the adult mammalian brain, Front. Cell. Neurosci. 7 (2013) 258.
    [27] K.C. Kondapalli, J.P. Llongueras, V. Capilla-González, H. Prasad, A. Hack, C. Smith, H. Guerrero-Cázares, A. Quiñones-Hinojosa, R. Rao, A leak pathway for luminal protons in endosomes drives oncogenic signalling in glioblastoma, Nat. Commun. 6
    M. Kirsch, C. Ikonomidou, G. Schackert, A. Temme, RNA interference targeting survivin exerts antitumoral effects in vitro and in established glioma xenografts in vivo, Neuro Oncol. 13 (10) (2011) 1074–1089.