Involvement of the JNK/FOXO3a/Bim Pathway in Neuronal Apoptosis after Hypoxic-Ischemic Brain Damage in Neonatal Rats
Authors: Jinlin Wu
Li Zhang
Jun Tang
Tao Xiong
Jinhui Li
Yi Qu
Deyuan Li
Xihong Li
Dezhi Mu
Journal: PloS one (2015)
Discipline: Multidisciplinary
Keywords: JNK; FOXO3a; neuronal apoptosis; hypoxia-ischemia; Bim
Affiliations: Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu 610041, China; Key Laboratory of Obstetric & Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, Sichuan University, Chengdu, Sichuan 610041, PR China., Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu 610041, China; Key Laboratory of Obstetric & Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, Sichuan University, Chengdu, Sichuan 610041, PR China; Department of Neurology, University of California San Francisco, San Francisco, CA 94143, United States of America.

Abstract:  c-Jun N-terminal kinase (JNK) plays a key role in the regulation of neuronal apoptosis. Previous studies have revealed that forkhead transcription factor (FOXO3a) is a critical effector of JNK-mediated tumor suppression. However, it is not clear whether the JNK/FOXO3a pathway is involved in neuronal apoptosis in the developing rat brain after hypoxia-ischemia (HI). In this study, we generated an HI model using postnatal day 7 rats. Fluorescence immunolabeling and Western blot assays were used to detect the distribution and expression of total and phosphorylated JNK and FOXO3a and the pro-apoptotic proteins Bim and CC3. We found that JNK phosphorylation was accompanied by FOXO3a dephosphorylation, which induced FOXO3a translocation into the nucleus, resulting in the upregulation of levels of Bim and CC3 proteins. Furthermore, we found that JNK inhibition by AS601245, a specific JNK inhibitor, significantly increased FOXO3a phosphorylation, which attenuated FOXO3a translocation into the nucleus after HI. Moreover, JNK inhibition downregulated levels of Bim and CC3 proteins, attenuated neuronal apoptosis and reduced brain infarct volume in the developing rat brain. Our findings suggest that the JNK/FOXO3a/Bim pathway is involved in neuronal apoptosis in the developing rat brain after HI. Agents targeting JNK may offer promise for rescuing neurons from HI-induced damage. 

Retracted: Yes

PubPeer comments:
 Figure 2. * Red boxes: The GAPDH panel in Figure 2A appear to look similar the 180 degrees rotated GAPDH panel in Figure 2C. A narrow red circle brings attention to a small smudge visible in the middle of the blot. ![file] , 
 Figure 2 and 3. * Green boxes (three different shades): The GAPDH panel of Figure 2E, GAPDH panel of Figure 3A, Akt panel of Figure 3G and GAPDH panel of Figure 3G appear to look similar, albeit shifted and stretched differently. * Blue boxes: The p-FOXO3a panel of Figure 3C and the FOXO3a cytoplasm panel of Figure 3E appear to look similar. * Orange boxes: The GAPDH panel of Figure 3C, the p-Akt panel of Figure 3G, and the GAPDH panel of Figure 4A appear to look similar. ![file] , 
 #2 ![file] , 
 Dear Dr. Bik, Thank you very much for pointing out these mistakes. As of this paper, The first author made the figures and did not discuss the results with the other authors before submission. I have told the first author to check the figures carefully and will response to you as soon as possible. Sincerely, Dezhi Mu , 
 Dear Dr. Bik, Iu2019m the first author of the paper. Thank you very much for helping us to find out the mistakes in this paper. I have checked the original data and found that the figures I got from the collaborating units were wrong. I am very sorry for these mistakes because I did not discuss the results with the coauthors including the corresponding author before submitting the paper. I will contact the editor of the journal to supplement the experiments and resubmit the corrected figures. Thank you again. Sincerely, Deyuan Li , 
 #5  I have checked the original data and found that the figures I got from the collaborating units were wrong.  As an interested by-stander, I want to thank Deyuan Li (in addition to the corresponding author) for responding so promptly. I have to ask, though, who are these  collaborating units ? Do you mean your co-authors, or someone not listed in the Author Contributions paragraph? , 
 This article has a retraction notice https: journals.plos.org plosone article?id=10.1371 journal.pone.0233069  After this article [1] was published, concerns were raised about similarities between western blot data reported in Figs 1u20134. Specifically: The following panels appear similar: Fig 1A GAPDH, Fig 2E GAPDH, Fig 3A GAPDH, Fig 3G Akt, and Fig 3G GAPDH. Aspect ratios are different between the similar panels in some cases. The GAPDH panels appear similar in Fig 2A and 2C, when one is rotated 180 degrees, and Fig 2C GAPDH lanes 2u20135 appear similar to Fig 3C FOXO3a lanes 1u20134. Fig 3C p-FOXO3a and Fig 3E FOXO3a cytoplasm panels appear similar. Fig 3C GAPDH (rotated 180 degrees), Fig 3G p-Akt, and Fig 4A GAPDH panels appear similar. In Fig 4A, the Bim bands appear similar to the CC3 bands in the middle blot, when rotated 180 degrees. The above issues have potential implications for the reliability of quantitative data reported in the corresponding Relative Optical Density graphs. The authors confirmed the duplications and noted that the original data underlying the figures of concern are not available. Without the original raw data, the concerns about data reporting in the published figures cannot be resolved. The above issues call into question the reliability of the articleu2019s results and conclusions, as a result, the PLOS ONE Editors retract this article. DL, JL, and DM agreed with retraction. The other authors either could not be reached or did not respond directly. ,