Fig. 5
Cell surface antigen recognition ability of rGFP-PDL1scFv. Raw264.7 cells (2.0 × 106 cells/well) were stimulated with lipopolysaccharide (LPS) (1 or 10 µg/mL) and stained using PE anti-mouse PD-L1 antibody or purified rGFP-PDL1scFv. Then fluorescence was detected by flow cytometry (a-c) and fluorescence microscopy (d-g). Representative flow cytometry histograms of staining with PE anti-mouse CD274 (B7-H1, PD-L1)  antibody (a) or rGFP-PDL1scFv (b) are shown, respectively. Each white histogram indicates non-stimulated Raw264.7 cells. Gray histograms indicate LPS-stimulated Raw264.7 cells, and orange histograms indicate LPS-stimulated Raw264.7 cells. Black bars indicate the gated region used to calculate PD-L1-positive cells. c The graph shows the measurement results for the  gated positive cell ratio in the stimulated or non-stimulated Raw264.7 cells stained with rGFP-PDL1scFv. Data are the mean ± SD (n = 3), and each dot on the plot represents one experiment. Representative fluorescence microscopy images of Raw264.7 cells stimulated with LPS (f, g) or without LPS (d, e) and stained with DAPI and PE anti-mouse CD274 (B7-H1, PD-L1) antibody (d, f) or rGFP-PDL1scFv (e, g) are shown (blue: DAPI, red: PE, green: GFP). Scale bar = 20 μm