Figure 3
From: MT1-MMP regulates urothelial cell invasion via transcriptional regulation of Dickkopf-3
Expression of DKK3 following targeted suppression of MT1-MMP and effects of actinomycin D on DKK3 mRNA. (A) Quantitative RT-PCR of DKK3 mRNA following knockdown of MT1-MMP (Si) and scrambled control (SC) at 24 and 48 h expressed as a ratio of untreated controls. (B) Western blot analysis of DKK3 secreted in media following knockdown of MT1-MMP at 48 h compared to scrambled control (SC) transfection and wild-type EJ28 cells (C48). Actin control was used against the cell lysate, whereas amphiregulin control was used against the media. (C) Densitometric analysis of the western blot shown in (B). Image J software was used to calculate the integrated density of the inverted image and expressed as a ratio in comparison with wild-type EJ28 cells (C48) (D) Following the MT1-MMP knockdown and scrambled control, transfection cells were incubated with actinomycin (10 μg ml−1) for 0, 1, 2, 4 and 8 h. The DKK3 mRNA levels were analysed by qRT-PCR and normalised against the housekeeping genes SDH and GAPDH. NTC, negative control.
