Figure 3.

Metastasis‐associated lung adenocarcinoma transcript 1 (malat1) M5 regulated the expression of specificity protein 1 (SP1) and SP1 target genes. A, malat1 siRNA significantly downregulated SP1 in A549 and H1299 cells, whereas overexpression of malat1 M5 caused accumulation of SP1 determined by immunofluorescence staining. Scale bar, 10 μm. B, After 48 h of transfection with pcDNA‐M5, blank vector pcDNA, si‐malat1 or si‐control in A549 and H1299 cells, total RNAs and protein, respectively, were collected, then (B‐1) RT‐PCR and (B‐2) western blotting were carried out to detect the levels of SP1,vascular endothelial growth factor (VEGF) and urokinase‐type plasminogen activator receptor (uPAR). GAPDH was used as control. B‐3, Relative density of SP1, VEGF and uPAR was plotted (relative to corresponding control) normalized to GAPDH ± SD from 3 independent experiments. ***P < .01. C, Analysis of luciferase intensity in cells cotransfected with pcDNA‐M5, blank vector pcDNA, si‐malat1 or si‐control, then with SP1‐Luc and the Renilla luciferase reporter plasmid. After 24 h of transfection, cells were assayed using a Dual‐Luciferase Reporter Assay System kit (Promega, Madison, WI, USA). All results are from 3 independent experiments. Values are mean ± SD, ***P < .01 compared with values from cells transfected with control