Both siDAAM1 and miR-335 promote growth and invasion of human astrocytoma U87-MG cells. (A) miR-335 expression in U87-MG and HEB cells was detected by real-time qRT-PCR. (B) Cell viability was detected by MTT assay. (C) Effect of siDAAM1 or miR-335 transfection on colony formation. (D) Effect of siDAAM1 transfection on DAAM1 protein expression. U87-MG cells were transfected with 50 nM siDAAM1 for 72 h. (E) Morphologic alteration (up-panel) and phallotoxins stained actin rearrangement (down-panel). Cells treated with siDAAM1 or miR-335 adopted a stellate morphology and collapsed their actin stress fibers together with increased membrane ruffling. (F) Cell invasiveness was detected by transwell invasion assay. (G left-panel) Western blot analysis was used to detect DAAM1 protein expression in U87-MG cells 72 h after transfection with 100 nM miR-335 mimics. (G right-panel) Endogenous DAAM1 expression in U87-MG and HEB cells was detected by western blot. GFAP expression was used as a biomarker of normal astrocytes. Results represent the means ± SD for three repeats. (*, p < 0.05;**, p < 0.01;***, P < 0.001). Original magnification in (E up-panel; F), 200 ×; (E down-panel), 400 ×.