Figure 5.
Figure 5. Rab11 expression compensates for defective Cdc42 signaling during large particle internalization. (A) Rab11 expression rescues large particle uptake defect in Cdc42-depleted cells. Control (shScr) and Cdc42 (shCdc42)-depleted COS1 cells were transfected with constitutively active Rab11 (Rab11Q70L) or a control plasmid and challenged with 4.5-µm beads. Uptake was quantified microscopically. (B) Rab11 expression rescues large particle uptake defect in NWASP−/− MEFs. NWASP−/− and NWASP−/− MEFs, expressing Rab11Q70L, were challenged with 4.5-µm beads, and uptake was quantified. (C) Rab7 expression does not rescue large particle uptake defect in Cdc42-depleted cells. Experiment performed as in A, using constitutively active Rab7 (Rab7Q67L). (D) Rab7 inactivation does not affect large particle uptake efficiency. COS1 cells were transfected with a control plasmid or a dominant-negative Rab7 (Rab7T22N) and challenged with 4.5-µm beads, and uptake was quantified. (E) Rab11 inactivation leads to a size-dependent uptake defect. COS1 cells were transfected with a control plasmid, dominant-negative Rab11 (rab11S25N), or dominant-negative Rac1 (Rac1T17N). Rac1T17N was included as a negative control. These cells were then challenged with 1.5- and 4.5-µm beads, and uptake was quantified. Error bars represent SEM. **, P < 0.01; ***, P < 0.001.

Rab11 expression compensates for defective Cdc42 signaling during large particle internalization. (A) Rab11 expression rescues large particle uptake defect in Cdc42-depleted cells. Control (shScr) and Cdc42 (shCdc42)-depleted COS1 cells were transfected with constitutively active Rab11 (Rab11Q70L) or a control plasmid and challenged with 4.5-µm beads. Uptake was quantified microscopically. (B) Rab11 expression rescues large particle uptake defect in NWASP−/− MEFs. NWASP−/− and NWASP−/− MEFs, expressing Rab11Q70L, were challenged with 4.5-µm beads, and uptake was quantified. (C) Rab7 expression does not rescue large particle uptake defect in Cdc42-depleted cells. Experiment performed as in A, using constitutively active Rab7 (Rab7Q67L). (D) Rab7 inactivation does not affect large particle uptake efficiency. COS1 cells were transfected with a control plasmid or a dominant-negative Rab7 (Rab7T22N) and challenged with 4.5-µm beads, and uptake was quantified. (E) Rab11 inactivation leads to a size-dependent uptake defect. COS1 cells were transfected with a control plasmid, dominant-negative Rab11 (rab11S25N), or dominant-negative Rac1 (Rac1T17N). Rac1T17N was included as a negative control. These cells were then challenged with 1.5- and 4.5-µm beads, and uptake was quantified. Error bars represent SEM. **, P < 0.01; ***, P < 0.001.

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