Research Paper Volume 14, Issue 21 pp 8645—8660

Figure 3. USP7 regulates cyclin F protein levels. (A) HeLa cells treated with DMSO, MLN4924 (10 μM), or P22077 (25 μM) for the indicated hours were lysed, and immunoblotted as indicated. β-Actin was the loading control. Asterisks denote non-specific bands. Fold changes were calculated with densitometric values for cyclin F and cyclin A2 blots using β-Actin as loading control. (B) HCT116 cells were treated with DMSO or P22077 (25 μM) for 5 h. Where indicated, cells were treated with MG132 (10 μM) for 4 h prior to harvest. Cells were then lysed and immunoblotted as indicated. Asterisks denote non-specific bands. Fold changes were calculated with densitometric values for cyclin F and cyclin A2 blots using β-Actin as loading control. (C) HCT116 cells were treated with DMSO or cycloheximide (CHX; 50 μg/mL) for 75 min. Where indicated, bafilomycin A1 (BafA1; 100 nM) and/or MG132 (12.5 μM) were added along with CHX. Cells were lysed, and immunoblotted as indicated. Asterisks denote non-specific bands.