Inh mir nc

miR-26b mimics (5′-UUCAAGUAAUUCAGGAUAGGU-3′), miR-26b inhibitor (5′-ACCUAUCCUGAAUUACUUGAA-3′) and their respective negative controls miR-NC (5′-UUCUCCGAACGUGUCACGUUU-3′) and inh-miR-NC (5′-ACGUGACACGUUCGGAGAAUU-3′) were synthesized by Shanghai GenePharma Co., Ltd. Keap1 cDNA (GenePharma Co., Ltd.) was cloned into pCDNA3.1 vectors (Invitrogen; Thermo Fisher Scientific, Inc.) by Shanghai GenePharma Co., Ltd. in order to construct corresponding Keap1 expression vector. The sequence of small interfering RNA (siRNA) against Keap1 (siKeap1, 5′-GCGCCAAUGUUGACACGGA-3′) and the sequence of the scrambled siRNA (siNC, 5′-GUACCUGACUAGUCGCAGAUU-3′) were synthesized by Shanghai GenePharma Co., Ltd. CFs (5×10⁴/ml) were transfected with miR-26b mimics (20 nM) or miR-NC (20 nM), the miR-26b inhibitor (20 nM) or inh-miR-NC (20 nM) and 20 nM vectors or 50 nM siRNA. The transfection was conducted using Lipofectamine^® 2000 transfection reagent (Invitrogen; Thermo Fisher Scientific, Inc.). At 48-h post-transfection, the subsequent experiments were conducted.

The shRNA specific to HANR (shHANR) and its negative control (shNC), miR-29b mimics and its negative control (miR-NC), miR-29b inhibitor and its negative control (inh-miR-NC), were synthesized by GenePharma (Shanghai, China). For overexpression of HANR and ATG9A, the HANR and ATG9A cDNA were cloned into pCDNA3.1 vector by GenePharma, respectively. Transfection was conducted with Lipofectamine 2000 following the manufacturer’s instructions.

The short hairpin RNA (shRNAs) targeting circCTDP1 (shcircCTDP1; 5′-UCA AGA AUG CAG GCU CAA C-3′) with negative control (shNC; 5′-UCUCCG AUGCAG GCU CAA C-3′), miR-320b mimics (5′-AAA GCU GGG UUG AGA GGG CAA-3′) with negative control (miR-NC; 5′-AAU UCU CCG AAC GUG UCA CUU-3′) and miR-320b inhibitor (5′-UUG CCC UCU CAA CCC AGC UUU U-3′) with negative control (inh-miR-NC; 5′-CAG UAC UUU UGU GUA GUA CAA-3′) were synthesized by GenePharma (Shanghai, China). The full length of HOXA10 was subcloned into pcDNA3.1 to overexpress HOXA10 levels with empty pcDNA3.1 serving as control. The pcDNA3.1 vector was bought from GenePharma (Shanghai). Transfection of the cells with shcircCTDP1 (10 nM) or shNC (10 nM) and the miR-320b mimics (10 nM) or miR-NC (10 nM) and miR-320b inhibitor (10 nM) or inh-miR-NC (10 nM) was conducted with Lipofectamine 2000 transfection reagent (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. The efficiency of transfection was determined in each experiment using RT-qPCR 24 h post-transfection. All functional experiments were carried out 48 h post-transfection.