Figure 2

Different forms of FIP200 protein were expressed in cultured mammalian cells. (A) HEK293T cells were mock-transfected (None) and transfected with an expression vector containing HA-tagged FIP200. Cell lysates were analyzed by Western blotting using antibody against FIP200. (B) NIH3T3, MEF, and 293T cells were lyzed in EBC and SDS sample buffers and analyzed by Western blotting using antibodies against FIP200 and γ-tubulin. (C) NIH3T3 cells were treated with UV then incubated in complete medium, and lyzed in SDS sample buffer (input). Cell lysates in SDS sample buffer were diluted with EBC buffer, and FIP200 protein was immunoprecipitated with antibody against FIP200, treated with phosphatase, and visualized by Western blotting using antibody against FIP200. Cell lysates were also analyzed by Western blotting using antibodies against COP1 and γ-tubulin. The ratio between upper and lower bands of FIP200 is also shown at the bottom of the FIP200 blot.