Illumina HiSeq 2500 paired end sequencing; GSM1599706: macrophage (exposed to S. enterica LPS) 1; Mus musculus; RNA-Seq

macrophage (exposed to S. enterica LPS) 1

We present a detailed single cell analysis of the macrophage response to LPS from Salmonella enterica.  By combining single cell transcriptional analysis, fluorescently labeled, LPS-coated beads, and cytometry we are able to distinguish the responses of macrophages that have internalized LPS-coated beads and those that have not. Overall design: Analysis of 96 single macrophages that were either: left untreated, were exposed to but did not internalize uncoated beads, were exposed to and internalized uncoated beads, were exposed to but did not internalize LPS-coated beads, or were exposed to and did internalize LPS-coated beads.

Single cell analysis of macrophages exposed to beads coated with LPS from Salmonella enterica subsp. typhimurium

Submitted by Gene Expression Omnibus on 14-SEP-2015

Single cells were sorted into individual wells of a 96 well plate containing  5 µl TCL buffer supplemented with 1% 2-mercaptoethanol (Qiagen, Valencia, CA) according to fluorescent phenotype using FACS.  RNA was isolated using 2.2x RNAClean SPRI beads (Beckman Coulter Genomics, Danvers, MA) To prepare amplified cDNA, we used a slightly modified version of the manufacturer's protocol from the SMARTer Ultra Low RNA Kit ((Clontech, Mountain View, CA) (See linked articule for details).  Fragmentation and library constuction were were done using the Nextera XT kit (Illumina, San Diego, CA) according to the manufacturer’s instructions, with all quantities adjusted the ¼ of the recommended volumes.

Single cell analysis of macrophages exposed to beads coated with LPS from Salmonella enterica subsp. typhimurium

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