FIGURE 1.
c-FLIPL interacted with TIP49. A, Coomassie Brilliant Blue staining of GST, GST-FLIPL, and His6-TIP49 expression. B, GST pulldown assays. Purified GST and GST-FLIPL were incubated with His6-TIP49 and pulldown proteins were separated by SDS-PAGE. C, 293T cells were co-transfected with HA-FLIPL and FLAG-TIP49, or HA-FLIPL alone. Cell lysates were immunoprecipitated with anti-FLAG antibody and probed with anti-HA antibody. D, endogenous IP assay was performed in A549 cells with stable expression of c-FLIPL. Cells were immunoprecipitated with anti-TIP49 antibody and the precipitations were blotted with anti-FLIP antibodies. Normal IgG as a negative control. E, schematic diagram of c-FLIPL and its mutants. Deletion mutants were tagged with HA in the N terminus. F, truncations of HA-FLIPL: DED, Δp12, P20, and ΔDED were co-transfected with FLAG-TIP49, respectively. Cell lysates were immunoprecipitated with anti-FLAG antibody and Western blotting with anti-HA antibody. Co-IP pulldown showed DED and Δp12 mutants interacted with TIP49.