Additional file 1 of TRIM22 suppresses Zika virus replication by targeting NS1 and NS3 for proteasomal degradation

Additional file 1: Figure S1. ISGs are induced by ZIKV infection in A549 cells. a-c qRT-PCR analysis of MX1, IDO1 and RSAD2 mRNA in A549 cells stimulated with increasing dose of ZIKV (FSS13025) infection. Figure S2. TRIM22 inhibits ZIKV infection in vitro. a Vero cells in 24-well plate were transfected with increasing amount of pM01-TRIM22 plasmid, 12 h later, the cells were infected with ZIKV GZ01 at MOI = 0.01, IFA of ZIKV E protein was conducted at 24 hpi. Scale bar, 100 μm. b Western blot analysis of lysates from A549 cells and TRIM22−/− A549 cells. c-d SH-SY5Y cells were transfected with pM01-TRIM22 plasmid, 12 h later, the cells were infected with ZIKV GZ01 at MOI = 0.1, ZIKV loads in cell lysates were measured by qRT-PCR at 24 hpi (c) and 48 hpi (d). e Western blot analysis of lysates from SH-SY5Y cells which were transfected with either NC siRNA or TRIM22 siRNAs. f SH-SY5Y cells were transfected with NC siRNA or TIRM22 siRNAs, 24 h later, the cells were infected with ZIKV GZ01 at MOI = 0.01, ZIKV loads in cell lysates were measured by qRT-PCR at 48 hpi. Figure S3. Model of BiLC system and IFA used to test the interactions between TRIM22 and ZIKV proteins. a Work model of GlucN-TRIM22 and GlucC-ZIKV NS1 or NS3 in BiLC system. b IFA of A549 cells that were co-transfected with pHA-TRIM22 and pHis-NS1 or pHis-NS3 plasmids, stained with HA and His tag antibody, then imaged by fluorescence microscope. Scale bar, 10 μm. Figure S4. TRIM22 mediates the K48 ubiquitylation of ZIKV NS1 and NS3 proteins. Western blot analysis of total cell lysates or immunoprecipitated proteins from HEK293T cells co-transfected with pM01-TRIM22, pHis-NS1 or pHis-NS3 and pHA-K48 (a) or pHA-K63(b) plasmids and treated with MG132, as indicated. Figure S5. TRIM22 inhibits YFV infection in vitro. a TRIM22 was over expressed in A549 cells, the cells were subsequently infected with YFV at MOI = 0.01, 12 h post transfection, YFV load in culture supernatant was measured by qRT-PCR. b TRIM22−/−A549 cells were infected with YFV, the cell lysates were measured by qRT-PCR. qRT-PCR data are means ± SEM from three independent experiments. **P < 0.05, **P < 0.01 and ***P < 0.001 by Student’s t test. Table S1. The list of ISG, ZIKV, DENV and YFV primers used in this study