Control oligonucleotide

HeLa cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Lonza, cat. #12-604F, Walkersville, MD, USA) supplemented with 10% fetal bovine serum (FBS; Welgene, cat. #S001-01, Gyeongsan, Republic of Korea). For plasmid transfection, plasmid DNA (1 µg/35-mm dish) was mixed with Effectene reagent (Qiagen, cat. #301425, Valencia, CA, USA) and added to a dish in which cells had been seeded 12 h before. The cell medium was replaced with fresh complete medium after 6 h. siRNA oligonucleotides against human MTFMT (5′-CCACAAACAGUCACAAAUATT-3′ and 5′-UAUUUGUGACUGUUUGUGGTT-3′), human ECSIT (5′-AUUGAUGUCAAACUCGUAGTT-3′ and 5′-CUACGAGUUUGACAUCAAUTT-3′), human TRAF6 (5′-GCGCUGUGCAAACUAUAUATT-3′ and 5′-UAUAUAGUUUGCACAGCGCTT-3′) and control oligonucleotides were purchased from GenePharma (Shanghai, China). For siRNA transfection, siRNA (87.5 pmol/35-mm dish) was mixed with Lipofectamine 2000 reagent (Invitrogen, cat. #11668019, Carlsbad, CA, USA) and added to a dish containing cells. The cell medium was replaced with fresh DMEM containing 5% FBS after 10–12 h.

SNP and diethylamine (DEA) NONOate were purchased from Sigma Aldrich (MO, USA). The RBM3 antibody (ab134946) was from Abcam (Cambridge, MA, USA), and antibodies against, AMPKα (#5832), p-AMPKα (#2535), AKT (#9272), p-AKT (#4060), p-ERK1/2 (#4370), p38 (#9212), p-p38 (#4511), p-JNK1/2 (#4668), p-MKK3/MKK6 (#12280), cleaved PARP (#9541), and β-actin (#4970) were form Cell Signaling Technology (Beverly, MA, USA). Antibodies against ERK1 (sc-93), and JNK1 (sc-474) were from Santa Cruz Biotechnology (CA, USA). The 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) was purchased from Sangon (Shanghai, China). Annexin V-FITC cell apoptosis kit was from Beyotime (Jiangsu, China). Inhibitors SB203580 (p38 MAPK) and SP600125 (JNK), were from Sigma Aldrich. Inhibitors LY294002 (AKT) and U0126 (MEK) were from Cell Signaling Technology. The siRNAs specific to human Rbm3, human miR-143 mimics and antisense inhibitor, and their negative control oligonucleotides were purchased from GenePharma (Shanghai, China).

miR-21-1 or miR-181b-2 mimic (dsRNA oligonucleotides) and miR-21-1 or miR-181b-2 inhibitors (single stranded, chemically modified oligonucleotides), and control oligonucleotides were ordered from GenePharma (Shanghai, China). Sequences are as follows: miR-21-1 mimic, 5’-CAACAGCAGUCUGUAAGCUGGC-3’
(sense) and 5’-CAGCUUACAGACUGCUGUUGUU-3’ (antisense); miR-181b-2 mimic, 5’-AACAUUCAUUGCUGUCGCUGGG-3’
(sense) and 5’-CAGCGACAGCAAUGAAUGUUUU-3’ (antisense); negative control mimic, 5’-UUCUCCGAACGUGUCACGUTT-3’ (sense) and 5’-ACGUGACACGUUCGGAGAATT-3’ (antisense); miR-21-1 inhibitors, 5’-GCCAGCUUACAGACUGCUGUUG-3’ (chemically modified by 29-Ome) and miR-181b-2 inhibitors, 5’-CCCAGCGACAGCAAUGAAUGUU-3’ (chemically modified by 29-Ome) and negative control inhibitors, 5’-CAGUACUUUUGUGUAGUACAA- 3’. MBrC, EPC or HEK293 cells were transfected with 50-100 nM of each oligonucleotide.

miR-148 mimics (dsRNA oligonucleotides), miR-148 inhibitor (single-stranded oligonucleotides chemically modified by 2′-Ome) and control oligonucleotides were commercially synthesized by GenePharma (Shanghai, China). Nonspecific RNA mimics (mimics control) or inhibitors (inhibitors control) were transfected as negative controls, and they have similar chemical properties as the miR-148 mimics or miR-148 inhibitor, respectively. Their sequences are as follows: miR-148 mimics, sense: 5′-UCAGUGCAUUACAGAACUUUG-3′, antisense: 5′-AAGUUCUGUAAUGCACUGAUU-3′; mimics control, sense: 5′-UUCUCCGAACGUGUCACGUTT-3′, antisense: 5′-ACGUGACACGUUCGGAGAATT-3′; miR-148 inhibitor, 5′-CAAAGUUCUGUAAUGCACUGA-3′; and inhibitors control, 5′-CAGUACUUUUGUGUAGUACAA-3′. HEK293 cells or macrophages were transfected with 30–100 nM of each oligonucleotides 24 h before LPS stimulation using Lipofectamine 2000^TM (Invitrogen).

miR-15b mimics (dsRNA oligonucleotides), miR-15b inhibitors (single-stranded oligonucleotides), and control oligonucleotides were ordered from GenePharma (Shanghai, China). The sequences are as follows: miR-15b mimics were 5′-CGAACCAUUAUUUGCUGCUUUA-3′ (sense), 5′-AAGCAGCAAAUAAUGGUUCGUU-3′ (antisense); negative control mimics were 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense), 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); miR-15b inhibitors were 5′-TAAAGCAGCAAATAATGGTTCG-3′ (chemically modified by 2′-Ome); negative control inhibitors were 5′-CAGUACUUUUGUGUAGUACAA-3′.

miR-2187 mimics (dsRNA oligonucleotides), and control oligo nucleotides were commercially synthesized by GenePharma (Shanghai, China). Their sequences are as follows: miR-2187 mimics were 5′-UUACAGGCUAUGCUAAUCUGU-3′(sense), 5′-AGAUUAGCAUAGCCUGUAAUU-3′(antisense); negative control mimics were 5′-UUCUCCGAACGUGUCACGUTT-3′(sense), 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); miR-2187 inhibitor (ssRNA oligonucleotides chemically modified by 2′-Ome) were 5′-ACAGAUUAGCAUAGCCUGUAA-3′; inhibitors control were 5′-CAGUACUUUUGUGUAGUACAA-3′.