Extended Data Figure 10. The effect of SLC46A1 and SLC46A3 expression on CDN-induced reporter activation.

a-b, Enforced expression of SLC46A1 and SLC46A3 affects the responses of THP-1 cells to CDNs. Control THP-1 cells (transduced with empty expression vector) or SLC46A1-transduced THP-1 cells (a), or control THP-1 cells or SLC46A3-transduced cells (b) were stimulated with 2’3’-RR CDA (1.25 μg/ml), 2’3’-cGAMP (15 μg/ml) or hIFN-β (100 ng/ml). tdTomato reporter expression was measured by flow cytometry 18-22h after stimulation. c-d, SLC46A1 or SLC46A3 depletions had little or no effects on cellular responses to CDNs, and combining SLC46A1 or SLC46A3 depletions with SLC19A1 depletion, had no additional effect compared to SLC19A1-depletion by itself. THP-1 cells were transduced with non-targeting control CRISPRi gRNAs or SLC19A1-targeting CRISPRi gRNA in combination with a second control CRISPRi gRNA or SLC46A1-targeting CRISPRi gRNA in (c) or SLC46A3-targeting gRNA in (d). Cells were stimulated with 2’3’-RR CDA (1.67 μg/ml), 2’3’-cGAMP (10 μg/ml), or hIFN-β (100 ng/ml). tdTomato reporter expression was measured by flow cytometry 18-22h after stimulation. Combined data of three independent experiments. Statistical analysis was performed using unpaired two-tailed Student’s t tests (a-b), or one-way ANOVA followed by Tukey’s post-tests when comparing only the effects of depleting SLC46A1 (c) or SLC46A3 (d). Data are means ± SEM of n=3 independent replicates.