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RNA extraction, reverse transcription, and real-time polymerase chain reaction
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Total RNA was isolated from the cells using TRIzol reagent (Invitrogen) according to manufacturer’s instructions. One microgram of RNA was reverse transcribed into complementary DNA (cDNA) to be used for qPCR. qPCR was performed using SsoAdvanced SYBR Green supermix and CFX Connect RT System (BioRad) to examine the expression of DUSP16 with ACTIN as internal control using respective primer pairs (Supplementary Table 2).
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