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. 2016 Sep 22;10:218. doi: 10.3389/fncel.2016.00218

Figure 6.

Figure 6

Manipulation of AGAP1 in neuronal dendrites alters transferrin trafficking. (A) Micrographs of 3 DIV E18 rat hippocampal neurons co-expressing eGFP and control plasmid, scrambled oligos, knock-down with siRNA, and overexpression of WT AGAP-1. Red: Alexa-transferrin (Alexa-Tf); Green: Rab11. White box outlines a ROI enlarged beneath the micrograph. Right panels: transferrin uptake was determined after 30 min (0 min). Left panels: recycling was assessed after 45 min following incubation with unlabeled transferrin (45 min). Scale bar = 10 μm. (B) Quantification of percent colocalization within ROIs of Rab11 puncta with Alexa-Tf. Data were quantified from three independent experiments (n = 10). At the 0 min time point no significant difference was observed between the colocalization of Rab11 and Alexa-Tf in scrambled vs. siRNA for AGAP1 (p = 0.6) and control plasmid vs. overexpressed AGAP1 (p = 0.9). At the 45 min time point, colocalization of Rab11 and Alexa-Tf was significantly different in scrambled vs. siRNA for AGAP1 (p = 0.0002) and for control plasmid vs. overexpressed AGAP1 (p = 0.0002).