FIG. 4.

GLS1 silencing reduced NPC proliferation. Human NPCs were transfected by control siRNA or GLS1 siRNA, then maintained in growth media for 3 days. NPC proliferation was determined by immunolabeling the Ki67 of proliferating cells in culture. (A) Representative pictures of Ki67⁺ (green) cells in control siRNA- and siGLS1-transfected NPC culture were shown. Nestin (red) was used to label NPCs. Nuclei (blue) were labeled with DAPI. (B) The percentage of Ki67⁺ cells was determined by counting the number of Ki67⁺ cells over the number of DAPI⁺ cells in each microscope field. Images were acquired from a Nikon Eclipse TE2000E fluorescent microscope. Magnifications: 20× objective lens. Scale bar=50 μm. (C, D) KGA and GAC mRNA levels were confirmed to be reduced in NPCs transfected by siGLS1 kept in growth media. Data are shown as mean±SEM of 10 fields in each experimental group for the three donors. *P<0.05, **P<0.01, ***P<0.001 compared with the control siRNA group, n=3. Color images available online at www.liebertpub.com/scd