Fig. 4 | Nature Communications

Fig. 4

From: Distinct submembrane localisation compartmentalises cardiac NPR1 and NPR2 signalling to cGMP

Fig. 4

Fluorescence recovery after photobleaching (FRAP) analysis for various NPR1-EYFP and NPR2-EYFP constructs. HEK293A cells were transfected with receptor constructs described in Supplementary Figure 7 and imaged 24–30 h later with and without MβCD treatment (1 mM for 1 h at 37 °C). Fluorescence was measured and bleached in a rectangular cell membrane region of interest using 488 nm laser and confocal microscope as described in Methods. Representative fluorescence recovery curves (a, b, d, e) and their time constant analysis after monoexponential fit (c, f) are shown. a NPR1-EYFP receptor mobility is sensitive to MβCD treatment. b This effect is abolished when TM domain of NPR1 is replaced by that of NPR2 in the NPR1/2 construct. d NPR2-EYFP receptor mobility is not sensitive to MβCD treatment. e However, it is changed by MβCD in the NPR2/1 construct which contained TM domain derived from NPR1. The opposite direction of MβCD effect (as compared to wild-type NPR1, compare e and a) might indicate that TM domain is crucial but not the only factor which affects NPR1 protein mobility. Data represent mean ± s.e.m. n = 10 cells each. * p < 0.05; *** p < 0.001; n.s., not significant by one-way ANOVA

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