Figure 1.

Construction of CAR^LunX

(A) Representative SPR affinity response curve of LunX antigen protein and the gradient concentration of s-35-8 antibody from 9.375 nM to 300 nM. (B) CAR^LunX cells used in functional experiments (schematic). VH, variable heavy chain; VL, variable light chain and the LunX-antigen specific CAR design. (C) The structure of each part of CAR^LunX. (D) Analyses of the sequence of s-35-8, CD137, and CD247 from the constructed plasmid by PCR. (E) Measurement of the mRNA expression of s-35-8, CD137, and CD247 from PCDH-MSCV-CAR^LunX-EF1-EGFP or empty vector-transfected 293T cells by RT-PCR. M represents the 100-bp DNA ladder and 1, 2, and 3 represent three repeated samples. One of three independent experiments is shown. (F) Immunofluorescence staining for c-myc tag of PCDH-MSCV-CAR^LunX-EF1-EGFP or empty vector-transfected 293T cells to analyze the transmembrane structure of the CAR. Scale bar, 100 μm.