Gene Expression Analysis by qPCR

Subconfluent cells were treated with various conditions for 48h. Total RNA was isolated by using the NucleoZOL reagent (Takara Bio USA Inc.), and subjected to reverse transcription (RT) reactions as previously described [63 (link)–66 (link)]. The RT cDNA products were used as PCR templates. The primers for the genes of interest were designed by using Primer3 Plus program (Supplementary Table 1). TqPCR reactions were carried out by using SYBR Green-based Forget-Me-Not™ qPCR Master Mix (Biotium Inc., Hayward, CA) on a CFX-Connect unit (Bio-Rad Laboratories, Hercules, CA) as described [67 (link)–70 (link)]. Relative gene expression was normalized to GAPDH by using the 2^−∆∆Ct method. All qPCR reactions were done in triplicate.

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Huang L., Zhao L., Zhang J., He F., Wang H., Liu Q., Shi D., Ni N., Wagstaff W., Chen C., Reid R.R., Haydon R.C., Luu H.H., Shen L., He T.C, & Tang L. (2021). Antiparasitic mebendazole (MBZ) effectively overcomes cisplatin resistance in human ovarian cancer cells by inhibiting multiple cancer-associated signaling pathways. Aging (Albany NY), 13(13), 17407-17427.

Publication 2021

NucleoZOL reagent CFX-Connect unit

Cdna Cells Gapdh Gene expression Genes Primers Reverse transcription Sybr green

Corresponding Organization : University of Chicago Medical Center

Other organizations : First Affiliated Hospital of Chongqing Medical University, Chongqing Medical University, Children's Hospital of Chongqing Medical University, Second Xiangya Hospital of Central South University, Central South University, Union Hospital, Huazhong University of Science and Technology

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Variable analysis

independent variables

Various conditions

dependent variables

Gene expression

control variables

GAPDH (used for normalization)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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