Nanometer photometer

Aortic specimens were ground in liquid nitrogen, and RNAiso Plus reagent (Takara 9109, Shiga, Japan) was used to extract total RNAs, and the concentration and purity of total RNA were detected by a nanometer photometer (IMPLEN). After that, reverse transcription was performed using PrimeScript RT reagent Kit with gDNA Eraser (TaKaRa RR047A, Shiga, Japan). Real-time quantitative polymerase chain reaction (RT-qPCR) was conducted using TB Green^® Premix Ex Taq (TaKaRa RR420A, Shiga, Japan) on an ABI Q3 7500 Real-Time PCR System (ABI). ACTB was used as internal controls, and the relative expression level of the target gene was calculated as 2−ΔCt, where −ΔCt = (Ct, target gene − Ct, ACTB). Statistical analysis was conducted with GraphPad Primer 8.0 (GraphPad Software Inc., GraphPad Prism 8.0.1.2). Primer sequences used in the study were listed in Supplementary Table 3.
IL-1β and CLEC4E concentrations in the serum were measured using a commercial ELISA kit according to the manufacturer’s instructions (#KET6013, EliKine Human IL-1β ELISA Kit; Abbkine Scientific Co., Ltd., Wuhan, China; # EK3805, Human C-type lectin domain family 4 member E ELISA Kit; Sabbiotech, College Park, MD, United States).

According to the manufacturer’s instructions, total RNA was extracted from OGSCs by the Trizol method(Invitrogen,Germany), and the concentration and purity of total RNA were detected by a nanometer photometer(IMPLEN, Germany). The measured optical density was 1.8 < A260/A280 < 2.2.RNA expression was detected by 2% agarose gel electrophoresis(Bio-Rad, USA). Table 1 contains the list of primers used in this study.
Table 1

Sequences used for Reverse Transcription- PCR

Gene	Forward primer (5′-3′)	Reverse primer (5′-3′)
GAPDH	AACGGATTTGGCCGTATTGG	CATTCTCGGCCTTGACTGTG
MVH	GTGTATTATTGTAGCACCAACTCG	CACCCTTGTACTATCTGTCGAACT
Fragilis	CTGGTCCCTGTTCAATACACTCTT	CAGTCACATCACCCACCATCTT
OCT-4	AGCTGCTGAAGCAGAAGAGG	GGTTCTCATTGTTGTCGGCT
Stella	CAGTCTACGGAACCGCATTG	CTTGGGAAAGGCGCTTTGAA
Dazl	GCCCTGCAATCAGGAAACAA	GGTTGGAGGCTGCATGTAAG
c-Kit	AACAGGACCTCGGCTAACAA	ACTGGCATCAGAGTTGGACA
ZP3	CTCTCCAGTTCACGGTGGAT	CGACTTTGAGATGGCAGGTG
Scp3	GCCGCTGAGCAAACATCTAA	GGCTTCCCAGATTTCCCAGA
Figla	AGCAGGAAGCCCAGTAAAGT	GCTCCTCAGGGCTTTGTTTC