Autophagy is upregulated in ovarian endometriosis: a possible interplay with p53 and heme oxygenase-1 Giulia Allavena, M.S., Patrizia Carrarelli, Ph.D.,

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Autophagy is upregulated in ovarian endometriosis: a possible interplay with p53 and heme oxygenase-1 Giulia Allavena, M.S., Patrizia Carrarelli, Ph.D., Barbara Del Bello, Ph.D., Stefano Luisi, M.D., Ph.D., Felice Petraglia, M.D., Emilia Maellaro, Ph.D. Fertility and Sterility Volume 103, Issue 5, Pages 1244-1251.e1 (May 2015) DOI: 10.1016/j.fertnstert.2015.02.007 Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Biochemical markers of autophagy indicate that autophagy is upregulated in OMA. Densitometric quantification of LC3-II protein (A), ratio LC3-II/LC3-I (B), and SQSTM1 (p62) protein (C) evaluated by Western blot. Results of protein levels in OMA and eutopic endometria of affected women (EEOMA) are reported as fold-increase over normal endometria from healthy women (NE) (mean value set to 1). (D, E) Representative Western blots. ***P<.001, OMA vs. NE or EEOMA. Fertility and Sterility 2015 103, 1244-1251.e1DOI: (10.1016/j.fertnstert.2015.02.007) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Autophagy-related genes are upregulated in OMA. Messenger RNA levels of ATG14 (A), BECN1 (Beclin-1) (B), ATG7 (C), and LC3B (LC3) (D). Gene expression values of EEOMA and OMA samples, measured by quantitative real-time PCR, are expressed as fold-change over NE samples. *P<.05 and *** P<.001, OMA vs. NE or EEOMA. Fertility and Sterility 2015 103, 1244-1251.e1DOI: (10.1016/j.fertnstert.2015.02.007) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Beclin-1 and Atg7 proteins decrease in OMA. Densitometric quantification of Beclin-1 (A) and Atg7 (B) proteins evaluated by Western blot. Results of protein levels in EEOMA and OMA samples are reported as fold-increase over NE samples (mean value set to 1). (C, D) Representative Western blots. *P<.05, OMA vs. NE and EEOMA, or EEOMA vs. NE. Fertility and Sterility 2015 103, 1244-1251.e1DOI: (10.1016/j.fertnstert.2015.02.007) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

Figure 4 p53 protein is decreased and HO-1 is induced in OMA. Densitometric quantification of p53 (A), Bcl-2 (B), and HO-1 (C) proteins evaluated by Western blot. Results of protein levels in EEOMA and OMA samples are reported as fold-increase over NE samples (mean value set to 1). (D–F) Representative Western blots. **P<.01 and ***P<.001, OMA vs. NE or EEOMA. Fertility and Sterility 2015 103, 1244-1251.e1DOI: (10.1016/j.fertnstert.2015.02.007) Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions