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Fig. 28.1 To select the shRNA-induced TauT-deficient 293 cell line, taurine uptake was measured.
The TauT-deficient cells exhibited a significant decrease in taurine uptake ( a ), which is reflected by
reduced TauT staining ( b ). This in turn reflects expression of the TauT gene ( c ). Thus, these results
confirm the success of shRNA-induced TauT knockdown in the 293 cells
confirmed such results. These results suggested that expression of TauT was sup-
pressed in the 293 cells by TauT shRNA.
28.3.2
Knockdown of TauT Signi fi cantly Suppresses
the Growth Rate of 293 Cells
Previous studies have shown that a TauT-deficient mouse has less body mass than a
wild-type control mouse (Heller-Stilb et al. 2002 ; Ito et al. 2008 ) , suggesting that
TauT may play a role in cell proliferation. In this study, an equal number (1 × 10 4 ) of
control and TauT-deficient 293 cells were cultured and counted every 3 days after
seeding. The results showed that inhibition of TauT significantly reduced the growth
rate of 293 cells in comparison to control cells over the course of 9 days (Fig. 28.2 ) .
28.3.3
Decreases in TauT Expression Inhibit
Colony Formation and Wound Healing
To test whether RNA-mediated reductions in TauT levels could influence the
ability of 293 cells to form colonies in soft agar, an equal number (1 × 10 4 ) of
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