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Overexpression of LINC00261 inhibits non–small cell lung cancer cells progression by interacting with miR‐522‐3p and suppressing Wnt signaling
RNA extraction and PCR
Total RNA was drawn out utilizing TRIzol reagent(Invitrogen, Carlsbad) from cultured cells or collectedtissues. Then reverse transcription reaction was performed to transfer total RNA to complementary DNA byusing PrimeScript RT reagent Kit with gDNA Eraser Kit(Takara, Ohtsu, Japan) following the manufacturer’s
protocol. The primers were product by Sangon Biotech(Shanghai, China). Primer Sequences used were listed in
Table 1. The SYBR Premix Ex TaqII Kit (Takara) wasapplied to perform qRT‐PCR in an Applied Biosystems
7900 Fluorescent qPCR System (Applied Biosystems,Foster City, CA). Relative quantification of gene expres
sion was calculated using the ΔΔCT calculation method.
Western blot analysis
Protein levels were evaluated by Western blot using theprotocol described previously.12 Primary antibodies usedwere listed below: rabbit anti‐GAPDH (ab37168; Abcam),rabbit anti‐SFRP2 (ab137560; Abcam), rabbit anti‐β‐catenin(ab32572; Abcam), rabbit anti‐c‐Myc (ab32072; Abcam) andrabbit anti‐cyclin D1(#2978; Cell Signaling Technology).
感謝河南省醫學科(kē)學技術研(yán)究項(xiàng)目引用(yòng)文(wén)獻(xiàn)
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