Fig 1: Short interfering RNA (siRNA) knockdown of CD133 alters Daoy MB cell function.(A) Western blot analysis of CD133 expression 48 h after transfection with siCD133-1, siCD133-2, scramble siRNA, or no transfection. ß-actin served as a loading control (left). Quantification of relative percent CD133 expression, measured by the densitometry of western blot bands and normalized to ß-actin (Scramble set to 100%) (right). (B) Fold change of the number of adhered cells per image after transfecting with siCD133-1, siCD133-2 or scramble siRNA, then culturing in ACM or DMEM (control) (DMEM scramble set to 1). (C) Average number of invading cells per image, counted on the lower surface of Boyden chambers with ACM or DMEM in the lower well, after transfecting with siCD133-1, siCD133-2 or scramble siRNA. (D) Total number of neurospheres counted with diameter length between 75–100 µm or >100 µm. Cells were transfected with siCD133-1, siCD133-2 or scramble siRNA and then cultured in the respective media for 48 h. Quantified values expressed as mean +/- SD. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.