Computational protocol: AngII-induced glomerular mesangial cell proliferation inhibited by losartan via changes in intracellular calcium ion concentration

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Protocol publication

[…] Total RNA was extracted with the addition of Trizol reagent (Invitrogen, USA) into all groups. cDNA was synthesized using 1–2 μg of total RNA. The primers were designed using the Primer Express software (Primer Premier 5.0) based on sequences obtained with the GenBank accession numbers. The sequences of the primers and amplicon sizes are shown in Table . PCR amplification consisted of one cycle at 95 °C for 30 s, followed by 40 cycles at 95 °C for 5 s and 60 °C for 20 s. Agarose gel electrophoresis was employed to analyze PCR products. Optical density analysis with glyceraldehyde-3-phosphate dehydrogenase (GAPDH, Invitrogen, USA) serving as an internal reference was used to measure mRNA expression in all groups. […]

Pipeline specifications

Software tools Primer Express, Primer Premier
Application qPCR
Chemicals Calcium, Prednisone, Trypan Blue, Losartan