Fig. 2: Integration of type II molecule ectodomains into cellular biosensors.

a Scheme depicting 4-1BBL-ζ and wildtype 4-1BBL constructs. b JE6.1 TPR cells or JE6.1 TPR cells expressing 4-1BBL or 4-1BBL-ζ were stained with a 4-1BBL antibody and assessed by flow cytometry. c Scheme depicting interaction of 4-1BBL-ζ on biosensor cells with 4-1BB on stimulator cells. d, e JE6.1 TPR, JE6.1 TPR 4-1BBL and JE6.1 TPR 4-1BBL-ζ were co-cultured with K562 cells expressing a membrane-bound anti-CD3 antibody fragment or K562 cells expressing 4-1BB and analyzed for reporter gene expression. d Representative histograms showing reporter gene expression of biosensor cells stimulated with K562 4-1BB is shown (data presented as NFκB-eCFP fluorescent intensity (FI)) e Representative experiment, performed in triplicates. Data is presented with mean ± SD of gMFI NFκB-eCFP. Data of one repeat experiment is provided within Source Data file. f 5×10⁴ JE6.1 TPR 4-1BBL-ζ cells were co-cultured with indicated count of K562-41BB cells (representative experiment, performed in triplicates). Data is presented with mean ± SD of gMFI NFκB-eCFP. Data of one repeat experiment is provided within Source Data file. g JE6.1 TPR 4-1BBL-ζ biosensor cells were co-cultured with stimulator cells as indicated. K562 4-1BB stimulators were preincubated with the indicated concentration of 4-1BB antibodies (n = 1 experiment, performed in triplicates). Data is presented as individual replicates with means ± SD of gMFI NFκB-eCFP. Source data for this figure are provided as a Source Data file.