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. 2013 May 31;8(5):e64514. doi: 10.1371/journal.pone.0064514

Figure 5. Morphology of coated pits in CALM-depleted cells.

Figure 5

a, Electron micrographs of control and CALM-depleted cells, showing the enlarged, non-constricted pits that accumulate during a CALM knockdown. b, Mixture of control and CALM-depleted cells, double labeled for CALM and Dab2, another component of the endocytic clathrin coat. The brighter labeling of Dab2 in the CALM-depleted cells can be correlated with the enlarged coated pits seen by electron microscopy. c, Ability of wild-type and mutant CALM constructs to rescue the morphology of clathrin-coated pits. For each condition, Dab2 labeling was quantified by automated microscopy. CALM knockdown with oligo 5 caused a ∼60% increase in spot intensity. The three CALM constructs are resistant to oligo 5. Both the wild-type and the mutant constructs were able to rescue the morphological phenotype. This indicates that the change in coated pit morphology upon CALM knockdown is unrelated to the VAMP-sorting function of CALM.