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Figure 3


Fig. 3. Depletion of {alpha}-adaptin and µ2 subunits of AP-2 blocks endocytosis of TfnR. (A) HeLa cells were transfected with the indicated siRNA oligonucleotides. Whole-cell lysates were prepared and immunoblotted with antibodies against {alpha}-adaptin, µ2-subunit or TfnR. (B) Live control cells were incubated with Alexa-Fluor-546-conjugated Tfn, then subjected to immunofluorescence using anti-{alpha}-adaptin antibody. (C) Live {alpha}-adaptin-depleted cells were incubated with Alexa-Fluor-488–Tfn, then stained with antibodies against {alpha}-adaptin and TfnR. (D) µ2-depleted cells were incubated with Alexa-Fluor-546-labeled Tfn, fixed, and then subjected to indirect immunofluorescence using anti-{alpha}-adaptin antibody. In C and D, the depleted cells are in the upper left parts of the panels.





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