Figure 5.

Melatonin induces the ubiquitination of Bim_EL and other proteins. (A) Bim_EL ubiquitination increased in porcine adherent granulosa cells treated with 10⁻⁹ M melatonin in the presence of 5 μM MG132 for 3 h. Bim was immunoprecipitated, and Western blotting was performed using anti-Bim and ubiquitin (Ub) antibodies. The asterisk indicates ubiquitinated Bim_EL appearing as a smear of bands with higher molecular weights. The level of protein ubiquitination increased in porcine adherent granulosa cells (B) and porcine primary granulosa cells (C) treated with 10⁻⁹ M melatonin in the presence of MG132 (5 μM) for 3 h. Western blotting was performed with an anti-ubiquitin antibody. Data are representative of three independent experiments.