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| Gene Synthesis | GenScript Real-time PCR (TaqMan) Primer Design (https://www.genscript.com/ssl-bin/app/primer) was used to design qPCR primers sets for pfn1, pfn2, pfn3 and pfn4 isoforms and GAPDH. The qPCR analysis was performed in CFX96 Real-Time PCR (BioRad) using SYBR Green I mixture in a final volume of 15 μl. 1 μg of total RNA was used for cDNA synthesis which was diluted 6 × for transcriptional analyses of profilin isoforms (pfn1, pfn2, pfn3, pfn4) in prenatal, postnatal and adult brain samples by using qRT-PCR. For each 15 μl reaction mixture was comprises SYBR Green I mix (Life technologies, Thermo Fisher Scientific, USA), 0.75 μl forward primer (10 μM) and 0.75 μl reverse primers (10 μM), cDNA (1 μl) and RNase free water. GAPDH (housekeeping gene) was used as an internal control in all q-PCR experiments. | Get A Quote |
Profilin (pfn) is an essential actin binding, post synaptic protein. Among its multiple isoforms, two (pfn1 and pfn2) have been reported in various organs including brain. pfn3 has been documented in kidneys and testes only, whereas pfn4 exclusively in testes. In this study, we report the presence of pfn isoforms, pfn3 and pfn4 in Mus musculus brain in addition to pfn1 and pfn2. Nine developmental stages were grouped into three classes, group 1: E10.5, E14.5, and E18.5 (prenatal), group 2: P0.5, P3.5, P5.5, P10.5 (postnatal) and group 3: P20.5, P60.5 (adult). Remarkably, all four isoforms exhibited variable yet significant expression in all discerning developmental stages. pfn3 presented high ... More
