Gene Expression Analysis by qPCR in Clinical Kidney Transplantation

Eikmans, Michael; Anholts, J.; Claas, Frans H.J.

doi:10.1007/978-1-4939-0733-5_14

Cited by 4 publications

(2 citation statements)

References 18 publications

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“…The PCR protocol used was 95 °C for 5 min, 40 cycles at 95 °C for 10 s, 60 °C for 34 s followed by a melting curve step of 60 °C up to 95 °C. The relative expression of the Rrp15 gene was normalized to the control gene actin .…”

Section: Methodsmentioning

confidence: 99%

Rrp15 affects cell cycle, proliferation, and apoptosis in NIH3T3 cells

Ren

Chen

et al. 2016

FEBS Open Bio

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Riken 2810430M08 (hereinafter referred to as Rrp15) is a newly identified and reported gene from the mouse genome. In our previous work, we found that the gene had a relationship with the proliferation and activation of T cells. Rrp15 protein is highly homologous with RRP15 (budding yeast), which has an important role in ribosomal RNA processing. We explored the potential function of Rrp15 in apoptosis, cell proliferation, and its involvement with RNA in the nucleus. We constructed a knockdown of the Rrp15 gene in NIH3T3 cells and then performed real‐time PCR, western blotting, flow cytometry, and immunofluorescence to determine the function of the Rrp15 gene. Knockdown of the Rrp15 gene suppresses proliferation and induces apoptosis. We also found that the Rrp15 protein was normally distributed in the nucleus and bound to RNA or pre‐RNA in the nucleus. Additionally, Rrp15 altered the activity of the 20S proteasome. Rrp15 promotes proliferation and inhibits apoptosis in NIH3T3 cells and may have a relationship with RNA in the nucleus.

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Section: Methodsmentioning

confidence: 99%

Rrp15 affects cell cycle, proliferation, and apoptosis in NIH3T3 cells

Ren

Chen

et al. 2016

FEBS Open Bio

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“…RNA was extracted using NucleoSpin columns (Macherey-Nagel) and tested for integrity by gel electrophoresis. Protocols for cDNA synthesis and real-time PCR have been previously described (Eikmans et al, 2013(Eikmans et al, , 2014. Levels of mRNA transcripts for CD55, CD46 and CD59 were normalised to the geometric mean signal of reference genes GAPDH and ␤-actin.…”

Section: Rna Extraction and Quantitative Pcr Analysismentioning

confidence: 99%