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. Author manuscript; available in PMC: 2015 Mar 18.
Published in final edited form as: Nat Commun. 2014 Sep 18;5:4925. doi: 10.1038/ncomms5925

Fig 4. Light-mediated disruption of clathrin dependent endocytosis.

Fig 4

(a) Schematic indicating light-mediated disruption of CLC function. (b) COS-7 cells expressing CRY2oligmCh-CLC before and after blue light. Scale bar, 7.5 μm. (c) COS-7 cell expressing CRY2oligmCh-CLC (inside dashed line) after exposure to blue light is defective for uptake for Alexa 488-transferrin (green), while surrounding untransfected cells show efficient uptake. Scale bar, 7.5 μm (d) Quantification of transferrin uptake in untransfected (mock) vs CRY2olig-CLC expressing cells exposed to dark, blue light, or light/3 hrs dark. Data represents average and S.E.M., n=15. ** p < 0.01 (e) Quantification of clathrin-coated pit dynamics within cells expressing CRY2olig-mCh-CLC in dark or after blue light (200 ms pulse, 488 nm), compared with a DsRed-CLC control. Data represents average and S.E.M., n=3 cells (over 30 clusters/cell counted). **, p < 0.01 (f) TIRF images of COS-7 cells expressing CRY2olig-mCh-CLC, showing clathrin pits at the cell surface before and after blue light exposure (488 nm, 350 ms). Arrow indicates mobile clathrin pit undergoing endocytosis. (g) Kymograph of CRY2olig-mCh-CLC-labeled clathrin pit showing increase in fluorescence after light exposure.

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