Skip to main content
Fig. 3 | BMC Biotechnology

Fig. 3

From: Modular assembly of synthetic proteins that span the plasma membrane in mammalian cells

Fig. 3

An ancillary fragment, R-GECO, is cytoplasmic and responsive to an induced Ca2+ influx. Pictorial representation of receptor orientation and response (a). COS-7 cells transfected with the plasma membrane labelled Lyn-Ceru showed a matte-like appearance (b and c) while those transfected with the ER labelled STIM1-mRFP showed a web-like fluorescence distribution (e and f). TLP-V-TM-RGECO showed a matte-like fluorescence distribution similar to Lyn-Ceru (b and c). Merged images illustrate resultant co-localization (d and g). Upon stimulation with 10 μM ATP, the Ca2+ concentration rapidly increased to a peak within a few minutes and then gradually declined to its basal level within two to three minutes (h). ER: endoplasmic reticulum, PM: plasma membrane, ATP: adenosine triphosphate, TM: transmembrane domain TLR4, TLP: fusion of TM with signal peptidase cleavage site from human immunoglobulin K, V: Venus fluorescent protein, RGECO: red fluorescent genetically encoded Ca2+ indicator, LC: Lyn-Ceru, STIM1: stromal interaction molecule 1. Scale bars are 10 μm. Images are false colored: CFP, cyan; YFP, green; mRFP, red. All insets show zoomed regions (4x) of structures in dotted rectangles. All experiments were repeated at least 3 times

Back to article page