Cfx96 qpcr machine

Total RNA was extracted from the coronary artery tissues and cells using Trizol reagent (Life Technologies, Carlsbad, USA) following the manufacturer’s instructions and then reverse-transcribed to cDNA using PrimeScript RT reagent kit (Thermo Fisher Scientific). The quantitative PCR was performed by using the SYBR ® Premix Ex Taq™ reagent (BioTeke, Beijing, China) on the CFX96 qPCR machine (Invitrogen, Carlsbad, USA) with the following steps: 10 min at 95°C; 35 cycles of 15 s at 95°C, 20 s at 60°C and 15 s at 72°C. The levels of mRNAs were calculated by the 2
^−ΔΔCt method, and normalized to that of
GAPDH as an internal reference. The primer sequences used in this study are listed in the
Table 2.

Table 2 Sequences of primers used in RT-qPCR

Gene	Sequence (5′→3′)
PDCD4	F: AAGAAAGGTGGTGGCAGGAGG
	R: TGACTAGCCTTCCCCTCCAA
HO-1	F: CGACAGCATGTCCCAGGATT
	R: TCGCTCTATCTCCTCTTCCAGG
VCAM-1	F: GATACAACCGTCTTGGTCAGCCC
	R: ATTGCCACAAGCAGAAAGACA
ICAM-1	F: AAACGGGAGATGAATGGT
	R: TCTGGCGGTAATAGGTGTA
p47	F: TCCCAAGTGGTTTGACGG
	R: CCTCCTCTTTCTGGCTGTG
WWP2	F: GAGATGGACAACGAGAAG
	R: CTCCTCAATGGCATACAG
GAPDH	F: GGTCGGGCAGGAAAGAGGGC
	R: CTAATCTTCTCTGTATCGTTCC

Total RNA was extracted from PC tissues and cells, and tumor tissues obtained from nude mice by using TRIzol Reagent (Invitrogen). Subsequently, a total of 1 μg RNA was reversed transcribed to cDNA by PrimeScript RT reagent Kit (Thermo Fisher Scientific, Waltham, MA, USA). The quantitative analysis of RAB11A mRNA and NORAD were performed by the SYBR ® Premix Ex Taq™ reagent (TaKaRa) and GAPDH was used as an internal control. The relative expression of miR-30a-5p was analyzed by SYBR PrimeScript miRNA RT‐PCR Kit (Takara Biotechnology, Dalian, China) with U6 as the internal control. qRT-PCR analysis was performed by CFX96 qPCR machine (Invitrogen, Carlsbad, CA, USA) with the following steps: an initial denaturation step at 95 °C for 3 min, followed by denaturation at 94 °C for 15 s, annealing at 55 °C for 25 s and extension at 72 °C for 15 s for 35 cycles. The quantification was calculated by the 2^−ΔΔCt method after normalized by internal control. The specific primer sequences used in our study were provided in Additional file 1: Table S1.