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Updated: Feb 8, 2026

In vitro Methylation Assay to Study Protein Arginine Methylation
Published on: October 5, 2014
Bardoxolone methyl induces neuritogenesis in Neuro2a cells
Namrata Chaudhari1, Palaniyandi Ravanan1
1Apoptosis and Cell Survival Research Lab, Department of Biosciences, School of Biosciences and Technology, VIT University, Vellore, Tamil Nadu, India.
Background:
Bardoxolone methyl (RTA 402, CDDOMe) has been long known for its anti-inflammatory and exceptional cytotoxic activity. The biological responses to CDDOMe are truly dose dependent. And owing to the structural modifications introduced in its parent molecule oleanolic acid, CDDOMe is able to form reversible adducts with cellular proteins containing redox sensitive cysteine residues. This nature of CDDOMe makes it a multifunctional molecule targeting multiple signaling pathways. This study was initiated to study the response of Neuro2a, a mouse neuroblastoma cell line to CDDOMe.
Methods:
Neuro2a cells were treated with CDDOMe and all trans retinoic acid (ATRA) for 4days and observed for neurite outgrowth. The neurite length was estimated using ImageJ software (Neuron growth plugin). Cell viability was investigated using MTT dye reduction and trypan blue dye exclusion method. Gene expression of differentiation markers was analyzed using quantitative PCR. Cellular localization of Tuj1 and synaptophysin in differentiated Neuro2a cells was observed using immunofluorescence.
Results:
CDDOMe ceased proliferation and induced dramatic neurite outgrowth in Neuro2a cells. These morphological changes were accompanied by time dependent increase in the mRNA levels of tyrosine hydroxylase, neurofilament 200 and synaptophysin. Besides, cytoskeleton protein Tuj1 and the synaptic vesicle protein synaptophysin were also observed to be localized in the neurites induced by CDDOMe.
Conclusions:
These early shreds of evidence suggest that CDDOMe induces differentiation in Neuro2a cells at concentrations ranging from 0.2 to 0.4μM and indeed contributes the existing knowledge on CDDOMe induced activities in cells.
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