Figure 2

OTUB1 is expressed in CRC tissues and cell lines and promotes migration and invasion. (A) The relative expression level of OTUB1 was examined by real-time PCR in 24 CRC tissues (T) and their paired normal mucosal tissues (N). β-actin was used as an endogenous control (**P <0.01). (B) OTUB1 expression at the protein level was detected by Western blot analysis in 5 pairs of CRC tissues and paired normal mucosal tissues. β-actin was used as a loading control. The relative mRNA expression level of OTUB1 was examined by real-time PCR (C) and the protein level was detected by Western blot analysis (D) in 7 CRC cell lines and 1 normal colon epithelial cell line (FHC). β-actin was used as an endogenous control (**P <0.01). (E) SW480 cells were transfected with the OTUB1 expression plasmid or empty vector for 48 hours, and the expression of OTUB1 at the protein level was examined by Western blot in the OTUB1 overexpression group (SW480-OTUB1) and the control group (SW480-Control). (F) Representative images showing the migration and invasion of SW480-OTUB1 and SW480-Control cells are shown. The number of tumor cells is quantified on the right. All data are expressed as the means of three independent experiments (**P <0.01). (G) SW620 cells were transfected with siRNAs against OTUB1 for 48 hours, and the expression level of OTUB1 at the protein level was examined by Western blot. Representative images showing migration and invasion and the quantitative analysis are shown in (H). The data represent the means of three independent experiments, and the error bars represent the SD (**P <0.01).