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Fig. 9. In HeLaM cells stably expressing a CD8-CI-MPR chimera, suppression of SNX2 induces a subtle change in the rate of sorting to the TGN. (A) Images depicting the kinetics of delivery of a cell-surface-labelled CD8-CI-MPR chimera to the TGN in HeLaM cells. Bars, 20 µm. (B) HeLaM cells stably expressing a CD8-CI-MPR chimera were treated with control, SNX1-specific, SNX2-specific, or SNX1- and SNX2-specific siRNA and subjected to an anti-CD8 antibody uptake experiment as described in Materials and Methods. Briefly, CD8-CI-MPR at the cell surface were labelled with anti-CD8, and allowed to internalise for 2, 8, 16 or 24 minutes. Cells were fixed and the amount of anti-CD8 that had reached the TGN46-labelled TGN was quantified by immunofluoresence using LCSLite (Leica). Results are expressed ± the s.e.m. from three independent experiments (except the 2 minute time point, which is from two independent experiments). The key indicates the siRNA treatment. (C) Western blot indicating representative levels of SNX1 and SNX2 suppression.
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