Kr118

The total RNA of cucumber leaves was extracted according to the instructions of the Tiangen plant total RNA extraction kit (DP432, Tiangen, Beijing, China), the reverse transcription kit was used to synthesize cDNA (KR118, Tiangen, China), and the SuperReal fluorescent quantitative premix reagent enhanced version (SYBR Green, FP205, Tiangen, China) was used for quantitative analysis. The primer information is shown in Table 1.

Myocardial tissue (10 mg) was collected from every group and homogenized thoroughly. Then, the TRIzol reagent (TR201-50; Tianmo Biotech, China) was used for total RNA extraction. Thereafter, the concentration and purity of extracted RNA sample were determined and the reverse transcription commercial kit (KR118; Tiangen, Beijing, China) was employed for reverse transcribing the purified RNA into cDNA. This cDNA was used as a template, and the 2× SYBR Green Master Mix (B21203; Bimake, TX, USA) and the downstream and upstream primers of the target gene were sequentially added to the reaction mixture (20 µL), and amplified with the help of a CFX96 RT-PCR apparatus (Bio-Rad, USA). Primer sequences of internal reference and target genes are presented in Additional file 1: Table S1. Finally, the 2^−ΔΔCt method was employed for analyzing the quantitative data.

Total RNA was isolated from hBMSCs cells using Trizol reagent (DP424, Tiangen, Beijing, China). All quantitative real-time PCR assays were carried out using three technical replicates and three independent cDNA syntheses. Reverse transcription reaction using cDNA synthesis kit (KR118; Tiangen, Beijing , China), qRT-PCR was performed using a fluorescence quantitative PCR kit (QP002, Fulengen, Guangzhou, China) and gene-specific primers (Table 1). GAPDH was used as an internal reference. The relative gene expression was determined using the 2^−ΔΔCt method. The PCR reactions were performed in triplicates.