Journal: Cell Cycle
Article Title: Thrombospondin-1 counteracts the p97 inhibitor CB-5083 in colon carcinoma cells
Figure Lengend Snippet: The steady-state level of THBS1 protein is increased by CB-5083. ( A ) HCT116 cells were treated with the indicated concentrations of CB-5083, Bortezomib and MLN4924 for 8 h. The steady-state levels of THBS1 and indicated proteins were analyzed by immunoblotting. Actin was used as a loading control. ( B ) Relative abundance of THBS1 protein was quantified (N = 3, SD error). ( C ) Relative abundance of known biomarkers of p97 inhibition (HERP1, CHOP and ATF4) was quantified (N = 3, SD error). ( D ) Mean fold changes (N = 3, SD error) for CDT1 and p21 with 3 μM CB-5083, 1 μM Bortezomib and 3 μM MLN4924. ( E ) An immunofluorescence assay for THBS1 (green), Actin (red) and DAPI (blue) in HCT116 cells treated with or without 1.5 μM CB-5083 for 8 h (N = 3, SD error).
Article Snippet: Human THBS1 cloned into pCMV3-Flag-THBS1 plasmid was purchased from Sino Biological (catalog number HG10508-NF).
Techniques: Inhibition, Immunofluorescence