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. Author manuscript; available in PMC: 2017 Nov 1.
Published in final edited form as: Biochim Biophys Acta. 2016 Aug 3;1863(11):2624–2636. doi: 10.1016/j.bbamcr.2016.07.008

Figure 7. Effect of ouabain on association between NKA and AT1R in kidney proximal tubule cells.

Figure 7

Figure 7

Figure 7

Immortalized human proximal tubule cells (HKC11) were transfected with mCherry-tagged AT1R (red fluorescence) and/or GFP-tagged human NKA (green fluorescence). Cells were treated with ouabain (1 nM) for 15 min. Excess ouabain was washed with PBS and the cells were imaged for epifluorescence (A), sensitized FRET (B), or TIRF (D) microscopy before and after treatment with ouabain (1 nM) as described in Methods. Representative images are shown from 8 individual experiments are shown (n=8). C, Sensitized FRET after photo bleaching was calculated using SlideBook4 software as described in Methods. Each bar represents corrected FRET (FRETc) as mean±se from 8 individual experiments (n=8). In each experiment 20–30 cells were imaged and data from all the cells were pooled and considered as one data point.