Shmmp 2

Short hairpin RNA (shRNA) targeting SPRY2 (shSPRY2; 10 nM; 5′-CCUUACCAUUCCUCCACUUTT-3′), shRNA targeting MMP-2 (shMMP-2; 10 nM; 5′-GCUGACCUGGAAGAGAACATT-3′) and their negative control (shNC; 10 nM; 5′-UUCUCCGAACGUGUC-3′), miR-124 mimics (10 nM; 5′-AACAUUCAACGCUGUCGGUGAGU-3′), NC mimics (10 nM; 5′-UUCUCCGAACGUGUCACGUTT-3′), miR-124 inhibitor (10 nM; 5′-ACUCACCGACAGCGUUGAAUGUU-3′) and NC inhibitor (10 nM; 5′-CAGUACUUUUGUGUAGUACAA-3′) were purchased from Shanghai GenePharma Co., Ltd. Cell transfection was performed in SRA01/04 (1×10⁵ cells/well) using Lipofectamine^® 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.) for 48 h at 37°C according to the manufacturer's instructions. The full-length of SPRY2 and MMP-2 were subcloned into pcDNA3.1 (10 nM; Shanghai GenePharma Co., Ltd.) to overexpress SPRY2 and MMP-2 levels with empty pcDNA3.1 (10 nM; Shanghai GenePharma Co., Ltd.) serving as the control. The efficiency of transfection was determined in each experiment using reverse transcription-quantitative (RT-q)PCR 48 h post-transfection. Subsequent experiments were performed at 48 h post-transfection.