Mix x synthesis kit

CRC tissues and cells were lysed with TransZol (TransGen, Beijing, China), and total RNA was extracted using an RNAsimple kit (Tiangen, Beijing, China). After performing concentration measure, RNA was reversely transcribed into cDNA using a FastKing RT Kit (Tiangen) or MiX-x™ synthesis Kit (TaKaRa, Dalian, China). Following that, SuperReal PreMix Color (Tiangen) was performed to detect the expression levels of circASXL1, miR-1205, or GRIK3. Data were assessed with the 2^-∆∆Ct method with U6 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as references. The sequences of forward (F) and reverse (R) primers were listed in Table S1.

Ox-LDL-induced HBMECs were collected and lysed using TransZol (TransGen, Beijing, China). RNA was reversely transcribed into cDNA with a High-Capacity cDNA RT Kit (Thermo Fisher) or MiX-x™ synthesis Kit (TaKaRa, Dalian, China). To determine the expression of circ_CHFR, checkpoint with forkhead and ring finger domains (CHFR), miR-15a-5p, and EGFR, SuperReal PreMix (Tiangen, Beijing, China) was employed. Data were analyzed with the 2^−∆∆Ct method with U6 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as references. The sequences of forward and reverse primers were circ_CHFR 5′-CCCTCTGCAAGGAAGCCACG-3′ and 5′-TGCGCCGCCTGCCTTCTGTA-3′; CHFR 5′-CTCGTGTTGGGCTCGTGTC-3′ and 5′-GAGCAGGTTTCACAGGAGTCA-3′; miR-15a-5p 5′-CTCACGTAGCAGCACATAA-3′ and 5′-ACCTCAAGAACAGTATTTCCAGG-3′; EGFR 5′-GACGACAGGCCACCTCG-3′ and 5′-ATCGCTGCTCCCCGAAGA-3′; U6 5′-CTCGCTTCGGCAGCACATATACT-3′ and 5′-ACGCTTCACGAATTTGCGTGTC-3′; GAPDH 5′-AACGGATTTGGTCGTATTGGG-3′ and 5′-CGCTCCTGGAAGATGGTGAT-3′.