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Fig. 8. Suppression of SNX2 does not result in a detectable defect in the steady-state distribution of the CI-MPR. (A) HeLa cells were treated with control, SNX1-specific siRNA, SNX2-specific siRNA or jointly with SNX1- and SNX2-specific siRNA duplexes for 72 hours. Cells were fixed and stained against endogenous SNX1, endogenous SNX2 and endogenous CI-MPR. In cells subjected to RNAi against SNX2, the CI-MPR remains at a perinuclear structure that has been previously characterised as the TGN. In cells in which SNX1 and SNX2 have been suppressed, the CI-MPR undergoes a limited redistribution at steady state to peripheral structures. Bars, 20 µm. (B) HeLa cells were treated twice with control, SNX1-specific siRNA, SNX2-specific siRNA or jointly with SNX1- and SNX2-specific siRNA duplexes for two consecutive periods of 72 hours. At this time, cells were washed into cycloheximide (40 µg/ml)-containing media for 0, 4, 8 or 12 hours. Cells were lysed and the levels of endogenous CI-MPR present were resolved by western blotting with CI-MPR-specific antisera, followed by volume integration. Results are presented graphically from the averages of four independent experiments.
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