Quantitative gene expression analysis

Total RNA was extracted from pellets or cells in each culture condition with Trizol Reagent (New Industry) following the manufacturer’s protocol. Total RNA was quantified with the Nanodrop Spectrophotometer (ND-1000, Thermo), and the reverse transcription reaction was performed on 1000 ng of RNA as previously described [13 (link)]. Quantitative real-time polymerase chain reactions (PCR) were performed on a Pikoreal 96 PCR System (Thermo) following the manufacturer’s procedures. The expression of type I collagen (COL I), type II collagen (COL II), type X collagen (COL X), aggrecan (ACAN), and SOX9 were analyzed with qRT-PCR with the gene-specific primers listed in Additional file 1: Table. S1. The target genes of each sample were normalized to the values of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as internal control. Relative expression of each gene was expressed as fold changes by the 2^−ΔΔCt method. Five samples of each group were measured. Statistical significance was marked with different letters (P < 0.05).

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Ning T., Guo J., Zhang K., Li K., Zhang J., Yang Z, & Ge Z. (2019). Nanosecond pulsed electric fields enhanced chondrogenic potential of mesenchymal stem cells via JNK/CREB-STAT3 signaling pathway. Stem Cell Research & Therapy, 10, 45.

Publication 2019

Nanodrop Spectrophotometer Pikoreal 96 PCR System

Aggrecan Cells culture Expression gene Genes Glyceraldehyde 3 phosphate dehydrogenase Pellets Polymerase chain reactions Primers Quantitative real time polymerase chain reactions Reverse transcription Sox9 Trizol Type i collagen Type ii collagen Type x collagen

Corresponding Organization :

Other organizations : Center for Life Sciences, Peking University, National University of Singapore

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Variable analysis

independent variables

Culture conditions

dependent variables

Expression of type I collagen (COL I)
Expression of type II collagen (COL II)
Expression of type X collagen (COL X)
Expression of aggrecan (ACAN)
Expression of SOX9

control variables

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as internal control

controls

No explicit positive or negative controls mentioned

Annotations

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