Leishmania RNA Virus controls the severity of muco-cutaneous leishmaniasis. Mus musculus

Comparison of the transcriptome of bone marrow derived macrophages infected with either metastasizing (M+, Lg13) or non-metastasizing (M-, Lg17) parasites after 24 hours of infection. Overall design: 3 experimental conditions; M+ infected BMMf vs. uninfected BMMf, M- infected BMMf vs. uninfected BMMf, M+ infected BMMf vs. M- infected BMMf, BMMf from 3 biologically independant infection experiments were used where BMMf were infected for 24hrs at a ratio of 1 BMMf to 10 parasites. Total RNA was extracted reverse transcribed into cDNA with either a Cy3 or Cy5 fluorescent tag and hybridized on to mouse 17k chip. Slides were scanned and differential BMMf gene expression was determined above 1.5 fold change and p less than 0.05. 6 slides per biological independant experiment were used including the dye swap, with 18 slides used in total.

本数据集对比分析了感染24小时后，分别被转移型（M+, Lg13）与非转移型（M-, Lg17）寄生虫感染的骨髓来源巨噬细胞（bone marrow derived macrophages, BMMf）的转录组。整体实验设计包含3组对照：感染M+寄生虫的BMMf与未感染BMMf、感染M-寄生虫的BMMf与未感染BMMf、感染M+寄生虫的BMMf与感染M-寄生虫的BMMf。本实验采用3次独立生物学重复的BMMf样本，感染时细胞与寄生虫的接种比例为1:10，感染时长为24小时。提取总RNA后反转录为cDNA，分别用Cy3或Cy5荧光基团标记，随后在小鼠17k基因芯片上进行杂交。对芯片进行扫描后，筛选差异倍数≥1.5倍且p值<0.05的BMMf差异表达基因。每次独立生物学重复包含6张芯片（含荧光染料互换对照），实验总计使用18张芯片。