FIG. 7.

Rnf6 overexpression increases Inha messenger level. (A) Complex formation between the NMPI probe and nuclear protein extracts from total testis cells, primary-culture Sertoli cells, and one of the 45T-1(RNF6) clones. The probe was incubated with the protein extracts either with or without an excess (100-fold) of unlabeled NMPI oligonucleotide. In the indicated lanes, the binding reaction was performed in the presence of anti-Rnf6 antibodies (SE2896). The arrows correspond to the protein complexes bound to the NMPI probe, and the arrowhead indicates the position of the higher-molecular-weight complex in the presence of antibodies. (B) Northern blot analysis of Rnf6 expression in primary-culture Sertoli cells (lane 1), in the 45T-1 cell line (lane 2), in 45T-1 cells transfected with the empty vector (lane 3), and in three independent 45T-1(RNF6) stable transformants (lanes 4, 5, and 6). The same membrane was hybridized in succession with probes for the Rnf6 (top) and Hprt (bottom) mRNAs. (C) The same RNA preparations as in panel B were reverse transcribed with oligo(dT) primers and tested by PCR for the presence of Inha (top) and Hprt (bottom) messengers.