Effect of UBLCP1 knockdown on gene expression in HeLa S3 cells. Homo sapiens

Human UBLCP1 is a unique nuclear phosphatase which has both ubiquitin-like domain and CTD phosphatase-like domain. UBLCP1 interacts with proteasome subunit Rpn1 and suppresses the activity of proteasome, but its role in gene expression is unknown. To identify genes whose expression levels are affected by UBLCP1 knockdown in human cultured cells, we performed gene expression profiling by microarray analysis. Overall design: HeLa S3 cells were seeded on 6-well plates at 1.0 × 10^5 cell/well and cultured at 37°C 24 h before transfection. Cells were transfected with either negative control siRNA or siRNA specific to human UBLCP1 gene using Lipofectamine RNAi MAX (ThermoFisher) and further cultured for 72 h at 37°C.

人源UBLCP1是一种独特的核磷酸酶，同时兼具泛素样结构域（ubiquitin-like domain）与CTD磷酸酶样结构域（CTD phosphatase-like domain）。UBLCP1可与蛋白酶体亚基Rpn1相互结合并抑制蛋白酶体活性，但其在基因表达过程中的功能尚未明确。
为了鉴定人源培养细胞中经UBLCP1敲低后表达水平发生改变的基因，我们通过微阵列分析（microarray analysis）开展了基因表达谱检测。
实验设计：将HeLa S3细胞以每孔1.0×10^5个细胞的密度接种于6孔板中，于37℃条件下培养24小时后进行转染。分别使用Lipofectamine RNAi MAX（ThermoFisher）转染阴性对照小干扰RNA（small interfering RNA, siRNA）或靶向人源UBLCP1基因的特异性siRNA，随后于37℃继续培养72小时。