Sybr green 1 master mix

RNA was extracted using TRIzol reagent (Invitrogen), treated with DNase I (Thermo Scientific) and converted into cDNA using random hexamer primers and RevertAid M-MuLV reverse transcriptase (Thermo Scientific). cDNA was quantified by SYBR green I master mix (FOREGENE) and gene-specific primers (Supplementary Table S2) on the StepOnePlus System (Applied Biosystems). PCR amplification reactions were all run in triplicates for each cDNA sample. For comparison, RNA level was first normalized to GAPDH mRNA, and the relative RNA level was determined by setting controls as 1.

RNA was extracted using TRIzol reagent (Invitrogen), treated with DNase I (Thermo Fisher Scientific, Rockford, IL, USA) and converted into cDNA using random hexamer primers and RevertAid M-MuLV reverse transcriptase (Thermo Fisher Scientific). cDNA was quantified using SYBR green I master mix (Foregene, Chengdu, China) and gene-specific primers on the StepOnePlus System (Applied Biosystems, Foster City, CA, USA). PCR amplification reactions were all run in triplicates for each cDNA sample. For comparison, RNA level was first normalized to B2M mRNA, and the relative level of RNA was determined by setting controls as 1. The qPCR primers are listed in Table S1.

Total cellular RNA was isolated from HSFBs using the TRIzol reagent (Invitrogen), and then random hexamer primers and RevertAid M-MuLV reverse transcriptase (Thermo Fisher Scientific) converted the RNA into complementary DNA (cDNA). The cDNA was subjected to qRT-PCR using gene-specific primers, and the SYBR green I master mix (FOREGENE, Chengdu, China) on the StepOnePlus System (Applied Biosystems, Foster City, CA, USA) was run in triplicate. Last, the relative quantities of miRNA and mRNA were calculated using the 2^−ΔΔCT method after normalization to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) levels. The qRT-PCR primers are listed in table S1.