Rnaiso kit

Our quantitative real-time PCR (qRT-PCR) analysis was conducted as follows. Roots were sampled from WT and phyb mutant plants at 4 weeks old and immediately frozen in liquid nitrogen. After total RNAs were isolated using RNAiso kits (Takara Bio, Shiga, Japan), first-strand cDNA was synthesized with MMLV Reverse Transcriptase (Promega, WI, USA) and the oligo(dT) 15 primer. Synthesized cDNAs were amplified using a SYBR Premix Ex Taq (TaKaRa) before qRT-PCR was performed on a Rotor-Gene Q instrument system (Qiagen, Hiden, Germany). For normalizing the amplified transcripts, we used a primer pair for rice ubiquitin 5 (OsUbi5/Os01g22490; Jain et al., 2006 (link)). All primers for these analyses are summarized in Table S1.

Here, RNAiso kits (TaKaRa, Dalian, China) were utilized for extracting
the total RNA from SHF bulges of cashmere goats and its stem cells. For the
expression detection of circRNA-0100 and related gene mRNAs, random primers
were utilized to synthesize the first strand cDNA with an M-MuLV cDNA synthesis
kit (Sangon, Shanghai, China), whereas for the expression detection of
miRNAs, the first strand cDNA was synthesized by a One-Step PrimeScript
microRNA cDNA synthesis kit (TaKaRa, Dalian, China). The divergent primers
for detecting the expression of circRNA-0100 were designed using the
CircPrimer program (Zhong et al., 2018). The convergent primers for mRNA
detection were designed by the Premier Primer 5.0 program (Premier Biosoft
International, Palo Alto, CA, USA). A combined internal control consisting
of UBC, YWHAZ, and SDHA was utilized for normalizing the gene expression
level (Bai et al., 2014). The corresponding mature sequences of all detected
miRNAs were retrieved from the miRNAsong database
(https://www2.med.muni.cz/histology/miRNAsong/index.php, last access: 16 April 2021). A combined
internal control consisting of let-7d-5p, miR-26a-5p, and miR-15a-5p was
utilized for normalizing the miRNA expression level (Bai et al., 2016). All
primers are provided in Table 1 with the corresponding detailed
information.

Total RNA was extracted using the RNAiso kit (Takara, Dalian, China). RNA was treated with DNase (Solarbio, Beijing, China) before cDNA synthesis. RNA was reversely transcribed into cDNA by the miScript reverse transcription kit (Qiagen, Hilden, Germany) and M-MLV reverse transcriptase kit (Promega, Madison, WI, USA). PCR amplification was accomplished on the ABI7500 Real-Time PCR System (Applied Biosystems; Thermo Fisher Scientific, Inc., Foster City, CA, USA) with the SYBR Green PCR kit (TaKaRa, Dalian, China), with U6 and β-actin as internal controls. The relative expressions of miR-15b-5p and YAP1 mRNA were calculated with 2^−ΔΔCt method [20 (link)]. The primer sequences are displayed in Table 1.Table 1.

Primer Sequences for RT-qPCR

Name	Primer sequences
miR-15b-5p	Forward:5’-TCGGGTAGCACACATAATGG-3’
	Reverse:5’-GTGCAGGGTCCGAGGT-3’
U6	Forward:5’-CTCGCTTCGGAGCACA-3’
	Reverse:5’-AACGCTTCACGAATTTGCGT-3’
YAP1	Forward:5’-ACCCACAGCTCAGCATCTTC-3’
	Reverse:5’-GCTGTGACGTTCATCTGGGA-3’
β-actin	Forward:5’-CCCACACTGTGCCCATCTAC-3′
	Forward:5’-GGAACCGCTCATTGCCAATG-3′