Pgph1 neo vector

For silencing of SP1, siRNAs (siSP1-1, 5ʹ-GCCAATAGCTACTCAACTACT-3ʹ; siSP1-2, 5ʹ-GGGTATGTTCCTAAGGTCATT-3ʹ) were synthesized, and nonspecific siRNA was used as a control (siNC). For LINC00339 knockdown, siRNA (si-00339, sequence: 5ʹ-GAGCCAGAAGTTGTCCTACTA-3ʹ) was synthesized, and a scrambled siRNA (si-ctrl) was used as a negative control for si-00339. The miR-378a-3p mimic, inhibitor, and corresponding negative control (NC) were obtained from GenePharma (Shanghai, China). For LINC00339 overexpression, the full-length cDNA of LINC00339 was amplified and subcloned into the KpnI and BamHI sites of the pcDNA3.1 vector (Invitrogen). The pcDNA3.1 empty vector (pc-NC) was used as a negative control for the LINC00339 overexpression vector. The shRNA for LINC00339 depletion (5ʹ-GACAGCTCCTAGAGTATCTTAATTGAT-3ʹ) was synthesized and inserted into the pGPH1/Neo vector (GenePharma). To generate cells stably overexpressing or silencing LINC00339, HCT-116 cells were transfected with pc-LINC00339 or sh-LINC00339, and the transfected cells were selected with 2 g/mL puromycin. For MED19 overexpression, the full-length cDNA of MED19 was PCR amplified and constructed into the XbaI and XhoI sites of the pcDNA3.1 vector, and an empty vector was used as a control (pc-NC). Transfection was performed using Lipofectamine 3000 (Invitrogen).