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Osteoclast response to low extracellular sodium

ABSTRACT: RAW 264.7 murine osteoclastic cells were cultured with recombinant cytokines (RANKL and M-CSF) in culture medium with normal sodium (NS; Na=140 mmol/l) and low sodium and reduced sodium (Na=120 mmol.l) while maintaining normal ostmolality with the addition of mannitol. After 24 hours, total RNA was extracted using PureLink kit (Invitrogen, Carlsbad, CA) according to manufacturer’s instructions, and purified using the Rneasy Mini Kit (Invitrogen). 5 µg RNA from each sample was converted into biotin-labeled cDNA and hybridized to GeneChip Mouse Genome 430 2.0 arrays containing 45,101 probe sets corresponding to known genes and expressed sequence tags. Total RNA was extracted using PureLink kit (Invitrogen, Carlsbad, CA) according to manufacturer’s instructions, and purified using the Rneasy Mini Kit (Invitrogen). 5 µg RNA from each sample was converted into biotin-labeled cDNA and hybridized to GeneChip Mouse Genome 430 2.0 arrays containing 45,101 probe sets corresponding to known genes and expressed sequence tags.

ORGANISM(S): Mus musculus

PROVIDER: GSE128152 | GEO | 2022/08/09

REPOSITORIES: GEO

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Niemann-Pick type C fibroblast analysis

Project description:Primary skin fibroblasts from four Niemann-Pick type C patients homozygous for the I1061T mutation and four control individuals were cultured under identical conditions in DMEM containing 10% fetal bovine serum. Cells were harvested at 50-70% confluency. mRNA was isolated with the FastTrack 2.0 mRNA isolation kit according to the manufacturer's instructions (Invitrogen, Carlsbad, CA). A reference RNA comprised of 10 cell lines was used as the control for each hybridized sample (Stratagene, La Jolla, CA). Both sample and reference RNAs were amplified using the MessageAmp II aRNA amplification kit (Ambion, Austin, TX). Set of arrays that are part of repeated experiments Keywords: Biological Replicate

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Pimary human keratinocyte knockdown with deltaNp63 specific siRNA

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RNA-Seq in HEK293T human cells

Project description:RNA-Seq procedure was used for analysis of expression in HEK293T cells. Cells were separately lysed with Trizol reagent (Invotrogen). Total RNA was extracted from cell lysed with Trizol reagent and PureLink RNA Micro Kit (Invitrogen) according to manufacture instruction. RNA quality was checked with BioAnalyser and RNA 6000 Nano Kit (Agilent). Poly(A)+ RNA was purified with Dynabeads® mRNA Purification Kit (Ambion). Illumina library was prepared from poly(A)+RNA with NEBNextNEBNext® Ultra™II RNA Library Prep Kit for Illumina® (NEB) according to manual. Sequencing was performed on HiSeq1500 with 50 bp read length. At least 10 millions of reads were generated for each sample.

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Identification of new full length Arabidopsis genes - pilot arrays

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