Source of chloroquine-induced vacuoles

In a recent post, I was musing about why the SmartFlares weren’t in the swollen chloroquine-induced vacuoles. Out of interest more than anything, I did a quick experiment to see what would happen if I pre-labelled the endocytic pathway with one coloured dextran, then added in a second with the chloroquine.

2015-06-22-schemeHere’s what happened:

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Discussion Points from SmartFlare Group Meeting

I recently discussed the SmartFlare project at a joint Sée and Lévy Group Meeting, partially extolling the virtues of Open Science but mostly focussing on the data acquired so far and some of the proposed experiments.

By Andreas E. Neuhold[CC BY 3.0], via Wikimedia Commons
Regarding the SmartFlare project, the three big questions that formed the basis of the talk were:

  • Why do only some cells take up SmartFlares?
  • Where are the SmartFlares?
  • Why are the VEGF and Scrambled SmartFlares fluorescent in puncta?

Below are some of the interesting points that were made during the meeting. Replies at the time (as best as I can remember them) in green:

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Interpretation of chloroquine results

The last post highlighted some of the results of using chloroquine to investigate the localisation and functionality of the SmartFlares. This post will deal solely with the interpretation of those results.

2015-06-10-chloroquine

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Effect of Chloroquine on SmartFlare Distribution and Intensity

In order to investigate why the SmartFlares seem to be fluorescent in puncta within the cells, we designed an experiment to try to inhibit endosomal maturation, thus (hopefully) preventing SmartFlare processing and release of fluorescence.

2015-06-02-Endosomal_Maturation

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