Pcdna empty vector nc

Small interfering RNA (siRNA) negative control (si-NC), siRNA against circ_0000228-1
(si-circ_0000228-1), siRNA against circ_0000228-2 (si-circ_0000228-2), pcDNA empty vector
(NC), pcDNA-circ_0000228 (circ_0000228), mimics negative control (mimics
NC), miRNA inhibitors negative control (inhibitors NC), miR-337-3p
mimics, and miR-337-3p inhibitors were available from GenePharma
Co., Ltd (Cat No. MIN0000578, Shanghai, China).
HeLa and C33A cells were planted in 6-well plates (3×10⁵ cells/mL) (Cat No.
353046, BD Biosciences, Bedford, MA, USA) and cultured at 37°C with 5% CO₂ for
24 hours. Then, cells were transfected using Lipofectamine® 3000 (Cat No. L3000015,
Invitrogen, Carlsbad, CA, USA) according to the manufacture’s instruction. Quantitative
real-time polymerase chain reaction (qRT-PCR) was performed to detect the transfection
efficiency.

From the Chinese Center for Type Culture Collection (Wuhan, China), we bought human HCC cell lines (HepG2, HCCLM3, Huh7 and MHCC97H) and human-derived liver cell line (HL-7702). All cells were cultured in RPMI-1640 medium (Thermo Fisher Scientific, Carlsbad, MA, United States) with 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, United States) and 100 μg/mL streptomycin and 100 U/mL penicillin (Invitrogen, Carlsbad, CA, United States) at 37 °C in 5% CO₂. From GenePharma Co., Ltd. (Shanghai, China), we purchased pcDNA-GMEB1, pcDNA empty vector (NC), small interfering RNA (siRNA) negative control (si-NC), siRNAs against GMEB1 (si-GMEB1#1 and si-GMEB1#2), pcDNA-YAP1 and siRNA against YAP1 (si-YAP1). HepG2 and Huh7 cells were transfected employing Lipofectamine^® 3000 (Invitrogen, Carlsbad, CA, United States) under the supplier’s instructions. At 48 h after transfection, Western blot was conducted to verify the transfection efficiency.