Hiscript 2 q rt kit

Total RNA was isolated using Trizol (Life Technologies, Carlsbad, CA, USA). HiScript II Q RT Kit (Vazyme, Nanjing, China) was used for reverse transcription. NovoStart^® SYBR qPCRSuperMix Plus (Novoprotein, Shanghai, China) was used to quantify gene expression level from the obtained cDNA. The primers for detecting OPNt, OPNa, OPNb and OPNc are listed in Table 1. GAPDH was used as the loading reference.

Total RNA was extracted from cell by using Trizol reagent (Sigma; T9424). The quantity and quality of RNA were determined using a ScanDrop2 nano-volume spectrophotometer (Analytik Jena), and reversely transcribed based on the HiScript II Q RT kit (Vazyme; R223) according to the manufacturer’s instructions. Amplification and real-time detection were performed on a qTOWER3 G real-time PCR system (Analytik Jena) by using ChamQ Universal SYBR qPCR Master Mix (Vazyme; Q711) in 20 μL reaction. The relative expression levels of each targeted gene were normalized by subtracting the corresponding mouse β-actin threshold cycle (CT) values by using the ΔΔCT comparative method. Three biological replicates per group were used for qPCR. Primers were synthesized by Sangon Biotech (Shanghai, China). Sequences of all primers used are provided in Additional file 1: Table S3.

Total RNA was extracted from the mycelia of each sample using the TaKaRa RNAiso Reagent (TaKaRa Biotechnology Co., Dalian, China). Each RNA sample was reversely transcribed into cDNA with a HiScript II Q RT Kit (Vazyme, R223-01, Nanjing, China). The expression levels of each gene under different culture conditions (as shown in the figure legends) were determined by quantitative real-time PCR with the primers listed in Table S1. For each sample, the quantification of the expression of the FgACTIN gene was performed as a reference. The experiment was repeated independently three times.

Total RNA was isolated from cell by using Trizol reagent (Sigma-Aldrich, St. Louis, MO, USA). cDNA was reversely transcribed using the HiScript II Q RT kit (Vazyme Biotech, Nanjing, China). Quantitative real-time PCR (qRT-PCR) analysis was performed in a qTOWER3 G real-time PCR system (Analytik Jena) by using ChamQ Universal SYBR qPCR Master Mix (Vazyme) according to the manufacturer’s instructions. The relative expression levels of mRNA were normalized to the expression of β-actin by using the 2-ΔΔCT method. Primers were synthesized by Sangon Biotech (Shanghai, China). Sequences of all primers were as follows: ACTB F 5′-CCTGTACGCCAACACAGTGC-3′, R 5′-ATACTCCTGCTTGCTGATCC-3′; AdipoR1 F 5′-ACGTTGGAGGGTCATCCCATA-3′, R 5′-AAACAGCACGAAACCAAGCAG-3′; AdipoR2 F 5′-CCCTCTCTTACAAGCCCATCA-3′, R 5′-GAGCCAGTCTGGTAGTACATCA-3′.

Total RNA was isolated using Trizol (Life Technologies, Carlsbad, CA, USA). HiScript II Q RT Kit (Vazyme, Nanjing, China) was used for reverse transcription. NovoStart® SYBR qPCRSuperMix Plus (Novoprotein, Shanghai, China) was used to quantify gene expression level from the obtained cDNA. The primers for detecting are listed in Additional file 1: Table S1. GAPDH was used as the loading reference. The cDNA was determined using a Quantitative Real-time PCR (Archimed X6, Rocgene, Beijing, China).