Fig. 2: Performance of NIR-FbLAGs nanobodies co-expressed with GCaMP6s in HeLa cells. | Nature Communications

Fig. 2: Performance of NIR-FbLAGs nanobodies co-expressed with GCaMP6s in HeLa cells.

From: Destabilized near-infrared fluorescent nanobodies enable background-free targeting of GFP-based biosensors for imaging and manipulation

Fig. 2

a Fluorescence images of HeLa cells coexpressing NIR-FbLAG16 or NIR-FbLAG30 anti-GFP Nb together with mTagBFP2 (negative control), mEGFP (positive control), and GCaMP6s. b Left, contrast of GCaMP6s only (n = 14) and GCaMP6s coexpressed with either NIR-FbLAG16 (n = 11) or NIR-FbLAG30 (n = 11) after the addition of 5 μM ionomycin. Right, the contrast of NIR-FbLAG16 and NIR-FbLAG30 for the data presented in the left graph. Data are presented as mean values  ±  s.e.m. for n  =  3 transfection experiments. c Change in fluorescence intensity of the cells co-expressing GCaMP6s (green) and NIR-FbLAG16 (red) in response to 5 μM ionomycin. d Change in fluorescence intensity of the cells co-expressing GCaMP6s (green) and NIR-FbLAG30 (red) in response to 5 μM ionomycin. In c and d, data are shown for 3 cells, and the arrow indicates the time point when 5 μM ionomycin was added. For imaging of mTagBFP2, mEGFP or GCaMP6s and NIR-FbLAGs, 390/40 nm excitation and 460/40 nm emission, 480/40 nm excitation and 530/40 nm emission, and 605/40 nm excitation and 640LP nm emission filters were used, respectively. Scale bar, 40 μm. Source data are provided as a Source Data file.

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