Phosphate-buffered saline (PBS) with a pH range of 7.35-7.65, consisting of 8000 mg/l NaCl, 200 mg/l KCl, 1150 mg/l Na2HPO4, and 200 mg/l KH2PO4 was used in this study. Single-cell fluorescence analysis was performed using the Beckman Coulter CytoFLEX flow cytometer (Beckman Coulter), where GFP fluorescent intensity was detected with both the FITC and PE channels or RFP fluorescent intensity was detected with PC5.5-A channels. The recombinant cells carrying plasmids pHY-UTRs-GFP or pHY-UTRs-RFP were cultured for 24 hours, washed twice with PBS, and then diluted 1:3 with PBS before analyzing 10,000 cells per assay. To test the fluorescent intensity of three chimeric proteins, the culture was diluted by a factor of 1:9 using PBS after being washed off. Multi-mode microplate reader (SpectraMax iD3; Molecular Devices) analysis on black 96-well plates (Corning, USA) was utilized to determine relative fluorescence intensity and cell optical density. An excitation of 480 nm and an emission of 520 nm were used to evaluate eGFP relative fluorescence intensity while cell density was assessed at a wavelength of 600 nm. FlowJo VX software was employed for data processing.
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