Fig. 2
From: Development of a novel PIG-A gene mutation assay based on a GPI-anchored fluorescent protein sensor
Expression of GPI-anchored GFP/mCherry. a Lentiviruses expressing GPI-anchored GFP and mCherry (fluorescent GPI-sensors) were prepared, and TK6 cells were infected. The expression of the GPI-sensors in TK6 cells were confirmed using an inverted fluorescent microscope. GPI-GFP and GPI-mCherry were confirmed to be targeted to the cell membrane. Proaerolysin (PA) resistant cells were labeled as GPI(−) cells, and the expression of GPI-sensors were compared under the fluorescent microscipe with GPI(+) cells. b The expression of GPI-sensors in TK6 cells were also confirmed by using flow cytometry using an LSRFortessa (BSL2). Signal intensity was measured by area on the histograms. Four group of cells can be detected: GFP and mCherry double-negative cells, GFP- or mCherry-single positive cells, and double positive cells. GFP and mCherry double positive TK6 cells were sorted by FACSAria II (BSL2), and single cells were expanded as GPI(+) cells for following experiments. c GPI(−) clonal cells were isolated, and the PIG-A gene was sequenced and compared with wild-type