Induction of apoptotic cell death via accumulation of autophagosomes in rat granulosa cells  JongYeob Choi, Ph.D., MinWha Jo, M.S., EunYoung Lee, M.S., DooSeok Choi, M.D., Ph.D.  Fertility and Sterility  Volume 95, Issue 4, Pages 1482-1486 (March 2011) DOI: 10.1016/j.fertnstert.2010.06.006 Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) Representative immunofluorescence images of granulosa cells cultured in serum-starved conditions. LC3-II was associated with punctate structures in granulosa cells cultured in serum-starved conditions alone (SC, top), but LC3-I had a diffuse expression pattern in the serum-starved granulosa cells treated with 3-methyladenine (3-MA, middle). The presence of punctate structures of LC3-II progressively increased in the serum-starved granulosa cells treated with bafilomycine A1 (Baf A1, bottom). (B) Transmission electron microscopic images of granulosa cells cultured in SC (left), SC + 3-MA (middle) and SC + Baf A1 (right). Arrows indicate representative autophagic vacuoles. N = Nucleus. (C, D) The percentages of dead and apoptotic cells in cultured rat granulosa cells under SC with/without 3-MA or BAf A1. The bar graph represents the mean ± standard error of results from five replicate experiments. ∗Statistically significant difference, P<.05. (E) Representative immunoblots of bax, bcl-2, and cleaved caspase-9 and caspase-3 proteins. In parts C and D, the bar graph represents the mean ± standard error of results from five replicate experiments. ∗Statistically significant difference, P<.05. Fertility and Sterility 2011 95, 1482-1486DOI: (10.1016/j.fertnstert.2010.06.006) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) Representative immunofluorescence images of granulosa cells cultured in serum-starved conditions. LC3-II was associated with punctate structures in granulosa cells cultured in serum-starved conditions alone (SC, top), but LC3-I had a diffuse expression pattern in the serum-starved granulosa cells treated with 3-methyladenine (3-MA, middle). The presence of punctate structures of LC3-II progressively increased in the serum-starved granulosa cells treated with bafilomycine A1 (Baf A1, bottom). (B) Transmission electron microscopic images of granulosa cells cultured in SC (left), SC + 3-MA (middle) and SC + Baf A1 (right). Arrows indicate representative autophagic vacuoles. N = Nucleus. (C, D) The percentages of dead and apoptotic cells in cultured rat granulosa cells under SC with/without 3-MA or BAf A1. The bar graph represents the mean ± standard error of results from five replicate experiments. ∗Statistically significant difference, P<.05. (E) Representative immunoblots of bax, bcl-2, and cleaved caspase-9 and caspase-3 proteins. In parts C and D, the bar graph represents the mean ± standard error of results from five replicate experiments. ∗Statistically significant difference, P<.05. Fertility and Sterility 2011 95, 1482-1486DOI: (10.1016/j.fertnstert.2010.06.006) Copyright © 2011 American Society for Reproductive Medicine Terms and Conditions