Azithromycin (AZM) is a widely used antibiotic, with additional antiviral and anti-inflammatory properties that remain poorly understood. Although Zika virus (ZIKV) poses a significant threat to global health, there are currently no vaccines or effective therapeutics against it. Here, we report that AZM effectively suppresses ZIKV infection in vitro by targeting a late stage in the viral life cycle. In addition, AZM upregulates the expression of host type I and III interferons and several of their downstream interferon-stimulated genes in response to ZIKV infection. In particular, we found that AZM upregulated the expression of MDA5 and RIG-I (pathogen recognition receptors induced by ZIKV infection) and increased the levels of phosphorylated TBK1 and IRF3. Interestingly, AZM treatment upregulated the phosphorylation of TBK1 without inducing the phosphorylation of IRF3 by itself. These findings highlight the potential use of AZM as a broad antiviral agent to combat viral infection and to prevent devastating ZIKV-associated clinical outcomes, such as congenital microcephaly.

29Single-cell technologies are becoming increasingly widespread and have been 30 revolutionizing our understanding of cell identity, state, diversity and function. However, 31 current platforms can be slow to apply to large-scale studies and resource-limited 32 clinical arenas due to a variety of reasons including cost, infrastructure, sample quality and requirements. Here we report DNBelab C4 (C4), a negative pressure orchestrated, 1 portable and cost-effective device that enables high-throughput single-cell 2 transcriptional profiling. C4 system can efficiently allow discrimination of species-3 specific cells at high resolution and dissect tissue heterogeneity in different organs, 4 such as murine lung and cerebral cortex. Finally, we show that the C4 system is 5 comparable to existing platforms but has huge benefits in cost and portability and, as 6 such, it will be of great interest for the wider scientific community. 7 8

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