Quantitative Analysis of Gene Expression

Total mRNA was obtained from clinical fresh tissues or cultured cells using QIAZOL (Qiagen, Shanghai, China). And 1 μg extracted mRNA was used for Complementary DNA (cDNA) synthesis with random primers and Maxima First Strand cDNA Synthesis Kit (Takara Bio, Inc., Otsu, Japan). Real-time quantitative PCR (RT-qPCR) was performed according to the manufacturer’s instructions using the SYBR Green Master Mix (TAKARA) and LightCycler480II system (Roche). GAPDH used as the reference gene for NAP1L1 or HDGF or c-Jun; and the relative expression of RNAs was calculated using formula [30 (link)]. The primers were listed in the Supplementary Table 3.

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Chen Z., Xie Y., Luo H., Song Y., Que T., Hu R., Huang H., Luo K., Li C., Qin C., Zheng C., Fang W., Liu L., Long H, & Luo Q. (2021). NAP1L1 promotes proliferation and chemoresistance in glioma by inducing CCND1/CDK4/CDK6 expression through its interaction with HDGF and activation of c-Jun. Aging (Albany NY), 13(24), 26180-26200.

Publication 2021

QIAZOL Maxima First Strand cDNA Synthesis Kit SYBR Green Master Mix LightCycler480II system

C jun Cdna Cultured cells Gapdh Gene Mrna Primers Real time pcr Rnas expression Sybr green Synthesis Tissues

Corresponding Organization :

Other organizations : TCM-Intigrated Cancer Center of Southern Medical University, Southern Medical University, Affiliated Hospital of Youjiang Medical University for Nationalities, Nanfang Hospital

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Relative expression of RNAs (NAP1L1, HDGF, c-Jun)

control variables

GAPDH (used as reference gene)

controls

Positive control: None mentioned
Negative control: None mentioned

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