Extended Data Fig. 1: Structures of CDNs used in this study and gating strategy for the genome-wide CRISPRi screens. | Nature

Extended Data Fig. 1: Structures of CDNs used in this study and gating strategy for the genome-wide CRISPRi screens.

From: SLC19A1 transports immunoreactive cyclic dinucleotides

Extended Data Fig. 1

a, Structures of the CDNs used in this study. b, Representative gating strategy for flow-cytometry-based sorting of the CRISPRi library of reporter-expressing THP-1 cells stimulated with CDNs. Cells were gated on the basis of their forward scatter (FSC) and side scatter (SSC) using gate P1. The P1 cells were subsequently selected for co-expression of BFP (fluorescent marker for the CRISPRi gRNAs) and GFP (marker for the expression of the reporter construct) (gate P2). Gate P3 excluded cell doublets present among P2 cells. Gate P4 selected for the lowest 25% of cells with respect to tdTomato expression and gate P5 selected for the highest 25%. c, Representative flow cytometry dot plots showing tdTomato expression in unstimulated cells or in cells stimulated for 20 h with cells for 20 h with CDN (2′3′-RR CDA). Data are representative of n = 3 biological replicates.

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