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. 2014 Feb 7;289(12):8375–8389. doi: 10.1074/jbc.M114.552133

FIGURE 3.

FIGURE 3.

Stable transfection of EGFP-HAS3 and silencing Rab10 expression in MCF7 cells. A, MCF7 cells were transduced with lentiviral vectors carrying doxycycline-inducible EGFP-HAS3 and hygromycin resistance genes and maintained in selective growth media with 0.25 mg/ml hygromycin. The expression of EGFP-HAS3 was induced with 1 μg/ml doxycycline, and the cells with high EGFP expression were selected using FACS as indicated with the bar. B, the selected MCF7-EGFP-HAS3 cells with doxycycline-inducible EGFP-HAS3 expression showed a dose-dependent increase of HA secreted in the medium after 48 h. Data represent mean ± S.E. (error bars) of three independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (one-way ANOVA, Tukey's test). C, EGFP-Rab10 was transiently expressed in the cells treated with Rab10 siRNA, along with the controls, and the cell lysate was analyzed by Western blots with an anti-GFP antibody. The expression of Rab10 was markedly reduced, whereas Rab7 was not altered.