Kinetics of LPS induction in wt and Cop1-ko BMDMs

Bone marrow was extracted from mice that are COP1-wt Rosa26-CreERT2 or COP1-floxed Rosa26-CreERT2 BMDMs were obtained by culturing bone marrow precursors in media containing 20% of supernatant from L929 cells. At day 4 of differentiation 4-OHT was added at 1uM to induce deletion of COP1 in BMDMs derived from COP1-floxed mice. At day 7 of differentiation, BMDMs were treated with 100 ng/ml of LPS or not. BMDMs were directly harvested in lysis buffer (from Qiagen RNeasy mini kit) at different time points (0h, 2.5h, 2.5h, 4h, 6h, 9h and 13h) following LPS stimulation. Three BMDMs preparations per group: G1: BMDMs from COP1-wt mice (expressing the wt allele of COP1) CRE positive. G2: BMDMs from COP1-floxed mice (expressing the floxed allele of COP1) CRE positive RNA from BMDMs following LPS treatment at different time points from mice that are COP1-wt Rosa26-CreERT2 or COP1-floxed Rosa26-CreERT2.

本数据集的实验样本制备流程如下：从COP1野生型（COP1-wt）结合Rosa26-CreERT2工具小鼠，以及COP1条件性敲除（COP1-floxed）结合Rosa26-CreERT2工具小鼠体内提取骨髓；将骨髓前体细胞置于含20% L929细胞上清液的培养基中经体外培养，以获得骨髓源性巨噬细胞（Bone Marrow Derived Macrophages, BMDMs）。在细胞分化第4天，向COP1-floxed小鼠来源的BMDMs培养液中添加1μM的4-羟基他莫昔芬（4-OHT），以诱导COP1基因的条件性敲除。细胞分化第7天时，将各组BMDMs分别用100 ng/ml脂多糖（Lipopolysaccharide, LPS）处理或设置空白对照。在脂多糖刺激后的不同时间点（0h、2.5h、2.5h、4h、6h、9h和13h），使用来自Qiagen RNeasy迷你试剂盒的裂解缓冲液直接收集各组BMDMs。每组设置3份独立的BMDM制备样本：G1组：来源于COP1-wt且Cre重组酶阳性的小鼠的BMDMs（表达COP1野生型等位基因）；G2组：来源于COP1-floxed且Cre重组酶阳性的小鼠的BMDMs（表达COP1条件性敲除等位基因）。本数据集包含上述两种基因背景小鼠的BMDMs经脂多糖处理后不同时间点的总RNA样本。