Cfx connect

Briefly, total RNA was extracted from cell lysates or cytoplasm and nuclear extracts using TransZol Up (Transgen, Beijing, China). RNA was analyzed quantitatively using a NanoDrop (NanoDrop Technologies, Rockland, DE, USA). Total RNA (1 μg) was reverse transcribed into cDNA using a cDNA synthesis kit (Transgen, Beijing, China) according to the manufacturer’s instructions. RT-PCR was performed in a Bio-Rad CFX connect real-time system detector with Transgen SYBR Green Supermix. The reactions were analyzed using Bio-Rad CFX Maestro software (Version 4.1). The threshold cycles (CTs) were calculated, and the relative gene expression was analyzed after normalizing to GAPDH. Experimental-related primers were designed and produced by Takara (Maebashi, Japan). The primers were as follows:

TransStart Tip Green qPCR SuperMix (Transgen) and CFX Connect were used for qRT-PCR (Supplementary Table S6). The reference gene for qRT-PCR was the peach RPL13 (ribosomal protein L13) gene. The results were unified using the 2^−ΔΔt calculation method.

Briefly, total RNA was extracted from the cell lysates, cytoplasm extracts or nuclear extracts by TransZol Up (Transgen, Beijing, China). RNA was quantitatively analyzed using a Nanodrop (Nanodrop Technologies, Rockland, DE, USA). Total RNA (1 μg) was reverse transcribed into cDNA using a cDNA synthesis kit (Transgen, Beijing, China) according to the manufacturer’s instructions. RT–PCR was performed in a Bio-Rad CFX connect real-time system detector with Transgen SYBR Green Supermix. The reactions were analyzed using Bio-Rad CFX Maestro software (Version 4.1). The threshold cycles (CT) were calculated, and the relative gene expression was analyzed after normalizing to beta-actin. The experimental primers were designed and produced by Takara (Japan) and are listed in Supplementary Table 5.