MC3T3-E1 single cell qPCR results (Biomark, Fluidigm)

Fluidigm Biomark qPCR raw data was pre-processed using Fluidigm Real-Time PCR Analysis software. Failed assays were removed. The default quality threshold cutoff (0.65) was used to identify potential artifacts. The baseline correction was to linear, and the Ct threshold detection method was set to Auto (Detectors). The Fluidigm Singular Analysis Toolset v3.6.2 was used to remove outliers using the IdentifyOutliers() command. Cell types used for this study: Wildtype MC3T3-E1 at differentiation stages T0, T3, T10, T14 and T21, Satb2-mutant cell lines C8 at T0 and C9 and T0 and T14.本研究的Fluidigm Biomark定量聚合酶链式反应（qPCR）原始数据采用Fluidigm实时定量PCR分析软件进行预处理。首先剔除失效的检测反应，以默认质量阈值截点（0.65）识别潜在实验伪影；基线校正采用线性模式，循环阈值（Cycle threshold，Ct）检测方法设置为自动（检测器模式）。随后使用Fluidigm单样本分析工具集（Singular Analysis Toolset）v3.6.2的IdentifyOutliers()命令移除数据中的异常值。 本研究所使用的细胞类型如下： 处于分化阶段T0、T3、T10、T14及T21的野生型MC3T3-E1细胞，以及Satb2突变体细胞系C8（T0阶段）与C9（T0、T14阶段）。