Figure 2From: Lipid metabolism enzyme ACSVL3 supports glioblastoma stem cell maintenance and tumorigenicity ACSVL3 knockdown depleted GBM neurosphere cells expressing stem cell markers. A. ACSVL3 expression was knocked down with two ACSVL3 siRNAs: ACSVL3 si3 and ACSVL3 si4. Quantitative RT-PCR (qRT-PCR) analysis indicated that compared to scrambled siRNA transfected cells (con), ACSVL3 mRNA level decreased ∼55-60% following ACSVL3 siRNA transfection for 72 hours in HSR-GBM1B cells. 18S was used as an internal control for qRT-PCR. Columns, mean relative ratio of ACSVL3 to 18S RNA from triplicate determinations. B-D. GBM neurosphere cells (HSR-GBM1A and HSR-GBM1B) were transiently transfected with ACSVL3 siRNAs and cultured in serum-free medium. Neurosphere cells were dissociated and subjected to flow cytometry to detect CD133-expressing cells (B) and ALDH-expressing cells (D). Total cellular proteins from neurospheres were collected and subjected to immunoblotting analysis for CD133 expression (C). ACSVL3 down-regulation significantly decreased the percentage of neurosphere cells with CD133+ and ALDH+. *: P < 0.05; **: P < 0.01.Back to article page