Fig 7.

SPF45 overexpression inhibits proliferation and ErbB2 expression in a phosphorylation-dependent manner. (A) SKOV-3 cells were retrovirally transduced with either empty pQCXIP vector (Vec), Myc-SPF45, Myc-SPF45-TASA (TASA), or Myc-SPF45-TDSD (TDSD). Cell lysates were immunoblotted with antibodies to Myc and actin. (B) Lysates from SKOV-3 vector and Myc-SPF45 cells were immunoblotted for SPF45, showing Myc and endogenous SPF45 expression. N.S., nonspecific. (C) SKOV-3 stable cell lines were immunostained with anti-Myc antibody and DAPI stained. (D) SPF45 overexpression inhibits cell proliferation in a phosphorylation-dependent manner. Cell counting of SKOV-3 stable cell lines was performed over 5 days. NS, not significant. (E) mRNA was isolated from the cells in panel A and subjected to real-time qRT-PCR using validated primers to ErbB2. The results are a combination of results of three independent experiments. The level of expression in the vector control cells was set to 1 in each experiment. (F) Cell lysates from the SKOV-3 stable cell lines in panel A were run on a gel and immunoblotted with antibodies to ErbB2 and actin. (G) Cell lysates from the SKOV-3 stable cell lines in panel A were run on a gel and immunoblotted with antibodies to Ets family transcription factors and actin.