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. 2015 Dec 15;109(12):2511–2522. doi: 10.1016/j.bpj.2015.10.047

Figure 5.

Figure 5

Two-color superresolution imaging of AQP4 and Kir4.1 labeled with Alexa 647 and 546, respectively. (A) Wide-field fluorescence microscopy showing AQP4 and Kir4.1 immunofluorescence in U87-MG cells cotransfected with Kir4.1 and M1-AQP4 or M23-AQP4. (B) Confocal microscopy of AQP4 and Kir4.1 immunofluorescence in brain cortical sections of wild-type mouse. (C) Blinking of single Alexa 546-labeled secondary antibodies in buffer containing 100 mM MEA. (D) Two-color wide-field (left panels) and superresolution images (right panels show the indicated boxed regions) of Kir4.1 and AQP4 in cultured U87-MG cells transfected with M1- or M23-AQP4. (E) Wide-field images of AQP4 and Kir4.1 (left panel) and superresolution images (right panels) showing distribution of AQP4 and Kir4.1 in parenchymal and perivascular regions of cortical sections. (F) Cross-correlation function, c(r), calculated for Kir4.1 versus AQP4 localizations in U87-MG cells transfected with M1- or M23-AQP4. The cross-correlation was averaged from four-to-six regions of the same size as those in (D). (G) The cross-correlation calculated for Kir4.1 versus AQP4 localizations in perivascular and parenchymal regions of cortical sections. To see this figure in color, go online.