Decreased expression of a subset of TLR-dependent genes by heme-inducible Spi-C

To determine the functions of Spi-C in innate immune responses, we investigated the overall gene expression patterns in M-CSF-BMDMFs prepared from Spicflox/flox and Lyz2-cre; Spicflox/flox mice. M-CSF-BMDMFs were stimulated with or without LPS following heme treatment and used for RNA-seq analysis. Overall design: Control and Spic–/– BMDMF pretreated with 40 µM hemin for 18 h were stimulated with (designated 'CNT_4' and 'cKO_4', respectively) or without (designated 'CNT_0' and 'cKO_0', respectively) 100 ng/ml LPS for 4 h.为明确Spi-C在固有免疫应答中的功能，我们对源自Spic^flox/flox与Lyz2-cre; Spic^flox/flox小鼠的M-CSF诱导骨髓源性巨噬细胞（M-CSF-BMDMF）开展了全基因表达谱分析。在经血红素处理后，我们对上述细胞分别施以脂多糖（Lipopolysaccharide, LPS）刺激或不施加刺激，随后进行RNA测序（RNA-seq）分析。整体实验设计如下：经40 μM氯化血红素（hemin）预处理18小时的野生型对照与Spic条件性敲除（Spic^-/-）BMDMF，分别以100 ng/ml LPS刺激4小时（分别命名为CNT_4与cKO_4），或不施加LPS刺激（分别命名为CNT_0与cKO_0）。