Trizol buffer

On the sixth day, OVA solutions of different doses were added to the DCs taken from mouse bone marrow, and the concentration of OVA in each hole was 0, 10, 100, and 10,000μg /ml, respectively. Three samples were tested for each group, and the culture lasted for 24 hours. After the collection of suspension cells, the cells, in each group, were separated and resuscitated in 0.5ml Trizol buffer (Takara, Dalian, China). RNA-seq was performed (Wuhan Kangce Co., LTD., China). Trizol extraction is a reliable method for obtaining high-quality total RNA. RNA-seq library was constructed by Illumina TruSeq RNA Sample Prep Kit version 2 (Illumina, San Diego, CA) and RNA was sequenced by Illumina HiSEquation 2000 platform (Illumina). Intergroup analysis: 10-0 DCs (10 DCs group compare with 0 DCs group), 100-0 DCs (100 DCs group compare with 0 DCs group), 10000-0 DCs (1000 DCs group compare with 0 DCs group).

Total RNAs of thyroid cancer tissues and the treated BCPAP cells were extracted by using TRIzol buffer (#9109, Takara, Japan). cDNA was synthesized with the Reverse Transcription kit (Takara, Japan). qRT-PCR assay was carried out with BestarTM qPCR Master Mix (#2043, DBI Bioscience, China) on ABI7300 system. The sequence of primers used in this study is shown in Table 1. The relative expression levels were analyzed using the 2^−ΔΔCt method [27 (link)].Table 1

The sequences of primers for RT-qPCR assay

Gene	Primer sequences
GAPDH	Forward: 5′-TGTTCGTCATGGGTGTGAAC-3′ Reverse: 5′-ATGGCATGGACTGTGGTCAT-3′
hsa_circ_0011385	Forward: 5′-TGACAACAATGAGCCCTACA-3′ Reverse: 5′-TTTCCTTGGCACTATACTGG-3′
miR-361-3p	Forward:5′-ACACTCCAGCTGGGTCCCCCAGGTGTGATTCTG-3′ Reverse:5′-CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGAAATCAGA -3′
U6	Forward: 5′-CGCTTCACGAATTTGCGTGTCAT-3′ Reverse: 5′-GCTTCGGCAGCACATATACTAAAAT-3′