Fig 1: c-Jun Activation Mediated SNHG1-Promoted MMP2 Transcription in Human Basal MIBC Cells(A and B) MMP2 mRNA levels were evaluated in indicated transfectants using real-time PCR; GAPDH was used as an internal control. *p = 0.0085 in (A), and *p = 0.0073 in (B) (significant difference). (C and D) Indicated cells were transfected with MMP2 promoter-driven luciferase reporter together with pRL-TK. Transfectants were seeded into 96-well plates and then subjected to determine MMP2 promoter activity by measuring luciferase activity. pRL-TK was used as an internal control to normalize transfection efficiency. *p = 0.0088 in (C), and *p = 0.0081 in (D) (significant difference). (E) Indicated cells were seeded into 6-well plates. After synchronization, cells were treated with Act D for indicated times. Total RNA was then isolated and subjected to real-time PCR analysis of MMP2 mRNA levels; GAPDH was internal control. *p = 0.0095 (significant difference). (F) Indicated cell extracts underwent western blotting for determination of SP1, P50, p-P65, P65, p-c-Jun, and c-Jun expression. (G) TAM67 was stably transfected into U5637(SNHG1) cells, and the stable transfectants were then identified for c-Jun(D) (TAM67) expression and determination of MMP2; GAPDH was used as an internal control. (H) Indicated cells were extracted for total RNA with TRIzol. Real-time PCR was used to determine MMP2 mRNA expression; GAPDH was used as an internal control. Results shown are means ± SD from at least triplicate experiments. *p = 0.0005 (significant difference).