Siklf4

Transfection of siControl (200 nM, Santa Cruz), siCHOP (200 nM, Bioneer Corporation) and siKLF4 (200 nM, Santa Cruz) into the cells were performed using Lipofectamine 3000 (Invitrogen) according to the manufacturer’s instructions.

Small-interfering RNA (siRNA) targeting human KLF4 (si-KLF4) and a nonspecific siRNA (si-NS) were purchased from Santa Cruz Biotechnology. Transfection was performed using Lipofectamine™ reagent (Invitrogen) following the manufacturer's instructions. At 24 h following transfection, LX-2 cells were incubated with or without TNF-α (10 ng/mL). Cells were then harvested and used for RT-PCR and Western blotting assays.

The NSCLC cell lines (A549 and H1975) were cultured in F-12 K or RPMI-1640 medium supplemented with 10% FBS (Hyclone, USA) at 37 °C and 5% CO₂.
Negative control, miR-1915-3p mimic, anti-miR-1915-3p were synthesized by GenePharma (Shanghai, China). SiNC (cat. no. sc-37,007), siSET (cat. no. sc-43,856), siMETTL3 (cat. no. sc-92,172), siYTHDF2 (cat. no. sc-78,661) and siKLF4 (cat. no. sc-35,480) were purchased from Santa Cruz biotechnology (Dallas, Texas, USA). The sequences of these oligonucleotides were as follows: miR-NC: 5′-UUCUCCGAACGUGUCACGUTT-3′; miR-1915-3p mimic: 5′-CCCCAGGGCGACGCGGCGGG-3′; anti-miR-1915-3p: 5′-CCCGCCGCGUCGCCCUGGGG-3′; anti-miR-NC: 5′-CAGUACUUUUGUGUAGUACAA-3′.
For SET overexpression, SET were amplified from H1975 cDNA, and subcloned downstream of FLAG tag in the pcDNA3.1-flag vector. The primers used for the coding sequences of SET amplification were as follows: 5′-GCGCGAATTC (EcoR I) ATGTCGGCGCCGGCGGCCAA-3′ (forward primer) and 5′-GCGCCTCGAG (Xho I) TTAGTCATCTTCTCCTTCATCCTC-3′ (reverse primer). For human KLF4 overexpression, the plasmid FLAG-KLF4 was generated, and the primers were as follows: 5′-GCGCGAATTC (EcoR I) ACATTAATGAGGCAGCCACCTG-3′ (forward primer) and 5′-GCGCGGATCC (BamH I) CGGGGGATTTAAAAATGCCTC-3′ (reverse primer).
Transient transfection of A549 or H1975 cells were performed using Lipofectamine 2000 (Invitrogen, USA).