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. 2010 Sep 20;285(48):37293–37301. doi: 10.1074/jbc.M110.157081

FIGURE 5.

FIGURE 5.

Effect of HNK-1ST or TNC knockdown by siRNA. a, Western blot of NSCs treated with HNK-1ST, TNC, or negative control siRNAs. Samples were collected after 72 h of siRNA treatment and then subjected to Western blot analysis with anti-HNK-1, anti-TNC, and anti-β-actin antibodies. β-Actin was detected as a loading control. Expression levels of TNC (b) and HNK-1 (c) were measured by densitometric analysis using a scanning densitometer (n = 3). d, effects of knockdown of TNC or HNK-1ST by siRNA on proliferation of NSCs. NSCs transfected with HNK-1ST, TNC, or negative control siRNAs were cultured as neurospheres for 72 h (n = 4). The proliferation rates of neurosphere-forming cells were measured by WST-8 assay at 24, 48, 72 h after siRNA transfection. e, number of neurospheres formed by siRNA-transfected NSCs counted at 72 h after lipofection (n = 4). f, proliferation rates of neurosphere-forming cells in the presence of control IgG, anti-HNK-1 or anti-TNC antibodies (20 μg/ml) estimated by WST-8 assay at 72 h in vitro (n = 3). The anti-HNK-1 and anti-TNC antibodies inhibit the proliferation of NSCs.