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Cast
 Jithan Ramesh as Easwar
 Varun Sandesh as Kumaran
 Uday Kiran as Anand
 Sangeetha
 Manjula Raman
 Veda
 Madhu
 Aishwarya Rajesh
 Kuladi Sister
 Deepika as herself
References
External links
Category:2009 films
Category:Tamil-language films
Category:2000s romantic comedy films
Category:Indian films
Category:Indian romantic comedy films
Category:Tamil film scores by Ilaiyaraaja caspase-3 activity.
3.7. Transfection of OPC-Like Cells {#sec3dot7-ijms-19-00139}
-----------------------------------
OGD-treated OPC-like cells were transfected with siRNA or with plasmid encoding for EBP50 or EBP50-DN mutant. Transfections were performed using JetPei (Polyplus Transfection, New York, NY, USA), according to the manufacturer's instructions. Briefly, cells were incubated with JetPei in serum-free medium for 20 min at room temperature. The resulting JetPei solution was removed and 5 mL of complete medium was added. A control treatment of JetPei only (no cells) was used to determine the effect of JetPei transfection reagent alone.
3.8. Western Blot Analysis {#sec3dot8-ijms-19-00139}
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Cells were lysed in RIPA buffer, and nuclear extracts were obtained by nuclear extract kit (ab113474, Abcam, Cambridge, UK). Protein concentrations were determined by the Bio-Rad assay (Bio-Rad Laboratories, Milan, Italy). Ten µg of proteins from lysates and 10 µg of nuclear proteins were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride (PVDF) membranes. Membranes were incubated with primary antibodies overnight at 4 °C, then washed and incubated with a secondary antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA). The immunoblots were developed using enhanced chemiluminescence (ECL) (Sigma-Aldrich) detection system. Densitometric quantification was performed using 3125841983
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