FIG. 5.

Prohibitin prevents GnRH-induced cell proliferation and alters cell morphology. A) LβT2 cells were stably transfected with siRNA to knockdown PHB expression, and Western analysis used to confirm the degree of knockdown in two of the clones (siPHB1 and siPHB2). The degree of knockdown was quantified relative to the controls after normalization with levels of ACTB in the same samples. B) The same two clones, as well as nontransfected αT3–1 and LβT2 cells, were cultured, and some of the cells exposed to GnRH (10 nM, 16 h) before carrying out a BrdU assay to evaluate cell proliferation. BrdU incorporation is expressed as a ratio to that in untreated cells of the same type. Mean ± SEM; n = 6. Statistical analysis was as described in Figure 4. C) Wild-type αT3–1 and LβT2 cells, as well as the LβT2, PHB1, and PHB2 knockdown cells, were photographed at two magnifications showing their different morphologies.