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Fig. 2 | BMC Cancer

Fig. 2

From: Aspirin inhibits epithelial-to-mesenchymal transition and migration of oncogenic K-ras-expressing non-small cell lung carcinoma cells by down-regulating E-cadherin repressor Slug

Fig. 2

Oncogenic K-ras up-regulates Slug transcriptionally to induce migration in A549 cells. a mRNA (upper panel) and protein (lower panel) expression levels of E-cadherin and Slug in untransfected-/ MEK-siRNA-/ ERK-siRNA-transfected A549 cells as determined by RT-PCR and western blot analyses, respectively. The inset represents the immunoblot analysis for the transfection efficiency of MEK-siRNA and ERK-siRNA in A549 cells. b Phase contrast images depicting migration of untransfected (upper left panel), MEK-siRNA-(middle left panel) and ERK-siRNA-(lower left panel) transfected A549 cells at 24 h as determined by wound healing assay was quantified and represented graphically (right panel). c Schematic diagram representing different regions of SLUG promoter and Slug coding region, and the sequential order of primer sets (sets 1–5 for promoter set 1 in upper panel and primer sets 1–7 for promoter set 2 in lower panel) to identify p-Elk-1 binding regions on SLUG promoter by ChIP analysis. d Schematic representations and RT-PCR data showing p-Elk-1-occupied region on SLUG promoter 1 (−325 to −129; primer sets 3 and 4) and SLUG promoter 2 (−496 to −384 and −181 to −40; primer sets 1 and 5, respectively) in A549 cells as determined by ChIP analysis. e Graphical representation of quantified relative protein expression of p-Elk-1 in untransfected, DN-K-ras-cDNA/MEK-siRNA/ERK-siRNA transfected A549 cells (right pannel) as determined by western blot analysis of Elk-1 and p-Elk-1 (left pannel) (f) Chromatin from oncogenic K-ras expressing control A549 cells, control-cDNA-/ DN-K-ras-transfected A549 cells was immunoprecipitated with p-Elk-1 antibody. PCR amplification was performed for p-Elk-1 binding regions on the SLUG promoter 1 (upper panel) and SLUG promoter 2 (lower panel). α-Actin/ GAPDH served as loading controls. Data are presented as mean ± SEM or representative of three independent experiments

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