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. 2011 Aug 23;105(7):1002–1011. doi: 10.1038/bjc.2011.303

Figure 6.

Figure 6

GS-168AT2 induced depletion of both CD9 and CD151 from cell surface. Human EC were incubated with either vehicle or GS-168AT2, and collected at the indicated time and washed twice with vehicle. Cell were then either lysed and resolved by SDS–PAGE, or directly analysed for the amount of CD9, CD151 and CD81 at the cell surface by FACS using the respective antibodies of these tetraspanins. All the results of FACS analysis were presented as the mean fluorescence intensity relative to control (hEC incubated with vehicle) and represent the mean±s.e. of four separate experiments realised in triplicate. Statistical significance were calculated with two-tailed Student's t-test (*P<0.05). (A) WB of the cell lysate with an anti-CD9 mAb monitoring relatively stable amounts of CD9 and the apparition of proteolytic fragment (16 kDa) of CD9 recognised by the anti-CD9 mAb with time in the presence of GS-168AT2. (B) Analysis of the hEC by FACS using anti-CD9 mAb showing that GS-168AT2 induced time-dependent decrease in CD9 at the cell surface. (C) WB of the cell lysate with anti-CD151 or CD81 mAbs monitoring decreased amounts of CD151 with time in hEC incubated with GS-168AT2 relative to vehicle, while there was not significant change in the level of CD81. (D) Analysis of the hEC by FACS using anti-CD151 mAb showing that GS-168AT2 induced time-dependent decrease in CD151 at the cell surface. (E) Analysis of the hEC by FACS using anti-CD81 mAb showing that GS-168AT2 did not induced significant changes in the level of CD81 at the cell surface. Western blots presented in this figure are representative images of at least three independent experiments.