Dntps

Aspartame and methotrexate were purchased from Sigma chemical company, St. Louis, MO, USA. Nitric oxide assay kit was purchased from Biovision Inc, Milpitas, CA, USA. Taq-Polymerase, DNTPs from (Genet Bio Inc, Yuseong-gu Daejeon, Korea) China, reverse transcription (RT) enzyme kit from (Thermo Fisher Scientific Inc, MA, USA), and other molecular grade chemicals from Merck Bangalore, India. All other chemicals were of analar grade obtained from Sisco research Laboratory, Bombay, India.

To synthesis cDNA, Suprime Script RTase, Oligo-dT and dNTPs (GeNet Bio, Korea) were reversely transcribed according
to the manufacturer’s instructions. The cDNA samples then were stored at -20 °C until further processes. The fidelity
of synthesized cDNA was then confirmed by polymerase chain reaction (Ampliqon Taq DNA polymerase Master Mix RED kit, Denmark)
using GAPDH primers. The final products were loaded on 2% agarose gel for electrophoresis (Figure 1).
Cycling conditions were as follows: initial denaturation at 95 °C for 5 min, followed by 40 cycles of denaturation at 95 °C for 30 s, annealing at 58 °C for 30 s, extension at 72 °C for 30 s and final extension for 5 min at 72 °C.

Aspartame and Methotrexate were purchased from Sigma chemical company, St. Louis, MO, USA. Nitric oxide assay kit was purchased from Biovision, USA. Taq-Polymerase, DNTPs from Genet Bio, China, Reverse Transcriptase enzyme kit from Thermo scientific, USA, primary antibody was purchased from Sigma and Cell signalling, USA, secondary antibody was purchased from Merck and other molecular grade chemicals were from Merck Bangalore, India. All other chemicals were of analytical grade obtained from the Sisco research laboratory, Bombay, India.