Expression of miR-145 and Its Target Proteins Are Regulated by miR-29b in Differentiated Neurons
Authors: Tanisha Singh
Abhishek Jauhari
Sanjay Yadav
Journal: Molecular Neurobiology (2018)
Discipline: NEUROSCIENCE & BEHAVIOR
Keywords: KLF4; MicroRNAs; NANOG; Neuronal differentiation; OCT4
Affiliations: Developmental Toxicology Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR- Indian Institute of Toxicology Research (CSIR-IITR), Vishvigyan Bhawan, 31 Mahatma Gandhi Marg, Lucknow, Uttar Pradesh, -226001, India., Academy of Scientific and Innovative Research (AcSIR), CSIR- IITR Campus, New Delhi, India., Department of Biochemistry, School of Dental Sciences, Babu Banarasi Das University, Lucknow, Uttar Pradesh, India., Developmental Toxicology Laboratory, Systems Toxicology and Health Risk Assessment Group, CSIR- Indian Institute of Toxicology Research (CSIR-IITR), Vishvigyan Bhawan, 31 Mahatma Gandhi Marg, Lucknow, Uttar Pradesh, -226001, India. .

Abstract:  MicroRNAs (miRNAs) are emerging as the most potential regulator of neuronal development. Recent studies from our lab and elsewhere have demonstrated a direct role of miRNAs in regulating neuronal differentiation and synaptogenesis. MicroRNA-145, a miRNA identified to regulate pluripotency of stem cells, downregulates the protein levels of reprogramming transcription factors (RTFs) like OCT4, SOX2, and KLF4 (cell, 137,647-658,2009). Studies have shown that miR-145 is multifunctional and crucial for fate determination of neurons. In our recently published study, we have identified a set of miRNAs including miR-145 and miR-29b families differentially expressed in SH-SY5Y cells exposed sequentially with retinoic acid + brain-derived neurotrophic factor (RA+BDNF) for differentiation into mature neurons (Mol Neurobiol (2016) doi: https://doi.org/10.1007/s12035-016-0042-9 ). In the present study, we have identified the role of miR-29b in upregulation of miR-145, which is upregulated after exposure of RA+BDNF in a P53-dependent manner. In differentiating SH-SY5Y cells, expression of miR-29b downregulates expression of P85Œ±, a P53 inhibitor, which results in upregulation of miR-145 and downregulation of RTF proteins. Ectopic expression of miR-145 and miR-29b in amounts equivalent to their endogenous expression has induced G1 phase cell cycle arrest. In conclusion, our studies have identified miR-29b as an upstream regulator of miR-145 and targets its RTF genes during differentiation of SH-SY5Y cells. 

Retracted: No

PubPeer comments:
  Same plot used for different treatment. , 
 Thank you for pointing out the mistake but we have already contacted the journal about this error! It was copy and paste error during preparing figures. -Abhishek Jauhari , 
 #2 If it was only copy and paste error, then why percentage shown in different quadrant are different in both the plots. Could the author give some explanation. , 
  There is an unusual similarity between western blot protein bands of Fig 6a: undifferentiated (control) group and Fig 6b: undifferentiated 0 days (highlight in Blue boxes), and between bands of Fig 6a: RA+BDNF exposed differentiated SH-SY5Y cells and Fig 6b: differentiated (8 Day) rat cerebral cortex NSCs (highlight in Red boxes). Can authors post full western blot images? , 
 Dear Sorex Ornatus Kindly go through the supplementary material of manuscript. We have already provided all unedited full western blot images there. Thank You Regards Abhishek Jauhari , 
  The same blots were flipped and reused in the paper for figure 4c and for figure 7a, patterns of nonspecific marks and dots can be clearly seen in supplementary figures 10a 11b. , 
  The same blot was used for Fig 7a Fig 7b for KLF4 and in the provided full blots in Supplementary Figure 7: Ladder lane was flipped and superimposed on the same blot and KLF4 bands are trimmed. , 
  How is it possible that there is not even a microscopic difference between blot probed with two different antibodies NANOG and KLF4 in supplementary fig 9a and 9b (full blots of figure 6b)? ,