Gene Expression Analysis of Adipose-Derived Stem Cells

Total RNAs of ASCs were extracted with RNeasy Mini kit (QIAGEN, Hilden). Reverse transcription was performed using High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Darmstadt), as instructed. All probes for gene analysis were obtained from Applied Biosystems: ADIPOQ (#Hs00605917_m1), AURKA (#Hs01582072_m1), AURKB (#Hs00945858_g1), CCP110 (#Hs00206922_m1), PLK1 (#Hs00153444_m1), PLK4 (#Hs00179514_m1), KIF2A (#Hs00189636_m1), KIF24 (#Hs00950248_m1), HDAC6 (#Hs00195869_m1), SMO (#Hs01090242_m1), GLI1 (#Hs00171790_m1), NANOG (#Hs04260366_g1), PTCH1 (#Hs00181117_m1), RUNX2 (#Hs01047973_m1), KLF4 (#Hs00358836_m1), c-MYC (#Hs00153408_m1), KLF6 (#Hs00810569_m1), PPARγ (#Hs01115513_m1), LEPTIN (#Hs00174877_m1), OCT4 (#Hs04260367_gH), SOX2 (#Hs01053049_s1), and GAPDH (#Hs02758991_g1). Real-time PCR was performed with a StepOnePlus Real-time PCR System (Applied Biosystems). The data were analyzed using StepOne Software v.2.3 (Applied Biosystems) as described previously [11 (link)].

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Ritter A., Kreis N.N., Roth S., Friemel A., Jennewein L., Eichbaum C., Solbach C., Louwen F, & Yuan J. (2019). Restoration of primary cilia in obese adipose-derived mesenchymal stem cells by inhibiting Aurora A or extracellular signal-regulated kinase. Stem Cell Research & Therapy, 10, 255.

Publication 2019

RNeasy Mini kit High-Capacity cDNA Reverse Transcription Kit ADIPOQ AURKA KIF2A HDAC6 c-MYC GAPDH StepOnePlus Real-time PCR System StepOne Software v.2.3

Ascs Aurka C myc Cdna Gapdh Gene Klf4 Klf6 Leptin Oct4 Plk1 Pparγ Ptch1 Real time pcr Reverse transcription Rnas Runx2 Sox2

Corresponding Organization : Goethe University Frankfurt

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Variable analysis

independent variables

None mentioned

dependent variables

Expression levels of the following genes: ADIPOQ, AURKA, AURKB, CCP110, PLK1, PLK4, KIF2A, KIF24, HDAC6, SMO, GLI1, NANOG, PTCH1, RUNX2, KLF4, c-MYC, KLF6, PPARγ, LEPTIN, OCT4, SOX2

control variables

None mentioned

positive controls

None mentioned

negative controls

None mentioned

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