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. 2005 Jun 29;25(26):6156–6166. doi: 10.1523/JNEUROSCI.1017-05.2005

Figure 4.

Figure 4.

Subcellular colocalization of variant and wild-type BDNF with sortilin. A, Differentiated PC12 cells were transfected with N-terminal HA-tagged variant (Met) or wild-type BDNF (Val). Cells were fixed after 48 h and permeabilized. Colocalization of BDNF and sortilin was visualized by costaining with anti-HA and anti-sortilin antibodies. In addition, untransfected differentiated PC12 cells were fixed after 48 h and permeabilized. Colocalization of endogenous sortilin and SecII was visualized by costaining with anti-sortilin and anti-SecII antibodies. Representative epifluorescence images are shown. B, Hippocampal neurons were transfected with N-terminal HA-tagged variant (Met) or wild-type BDNF (Val) and Myc-tagged sortilin. Cells were fixed after 48 h and permeabilized. Colocalization of HA-BDNF and Myc-sortilin was visualized by confocal microscopy, as described in Materials and Methods, using epitope antibodies. All neurons were confirmed using MAP2 antibodies. In addition, untransfected neurons were fixed after 48 h and permeabilized. Colocalization of endogenous sortilin and SecII was visualized by costaining with anti-sortilin and anti-SecII antibodies. Representative epifluorescence images are shown. Scale bars, 20 μm.