Figure 6.

Generation and localization of coilp1 fusion proteins, and stability of endogenous coilp1. (A) Western blot of ectopically expressed GFP-coilp1 and myc-coilp1. IPs were conducted using either anti-GFP or anti-myc antibody and then detected using coilin polyclonal antibody. Normal Mouse IgG was used as a negative control. Input represents 1.5% of lysate used in IPs. (B) Immunofluorescence of coilp1 fusion proteins in order to assess co-localization with Cajal bodies by using SMN monoclonal antibody. (C) Immunofluorescence of coilp1 fusion proteins and co-staining with coilin antibody. (D) Cyclohexamide treatment of HeLa cells to determine the half-life of coilp1. At 10 hours, coilp1 is reduced 51.9% relative to coilin (n = 3 experimental sets, p = 1.9×10⁻⁴). At 16 hours, coilp1 is reduced 77.9% relative to coilin (n = 3 experimental sets, p = 5×10⁻⁴).