Virologica Sinica

Supplementary Data

Enterovirus D68 infection upregulates SOCS3 expression to inhibit JAK-STAT3 signaling and antagonize the innate interferon response of the host

Yuling Zhanga, 1, Leling Xua, 1, Zhe Zhanga, 2, Xin Sua, 2, Zhiyun Wangb, *, Tao Wanga, c, *

a. School of Life Sciences, Tianjin University, Tianjin, 300072, China
b. School of Environmental Science and Engineering, Tianjin University, Tianjin, 300072, China
c. Institute of Tianjin Key Laboratory of Function and Application of Biological Macromolecular Structures, Tianjin, 300072, China
1Yuling Zhang and Leling Xu contributed equally to this work.

*Corresponding authors.
wangtaobio@tju.edu.cn (Tao Wang);
ORCIDs: https://orcid.org/0000-0003-1490-7407;

zhiyun_wang@tju.edu.cn (Zhiyun Wang);
ORCIDs: https://orcid.org/0000-0001-5236-8948.

Supplementary Fig. S1. A–D HEK293T cells and RD cells were infected with EV-D68 (MOI = 0.1) at
various times (6, 12, 24 h), and then the cells were collected. EV-D68-infected HEK293T cells and RD cells at different MOI (0.01, 0.1, and 1) for 24 h, and then cells were collected. The protein levels of STAT1 and p-STAT1 Y701 were measured by western blot. E-H HEK293T cells and RD cells were infected with EVD68 (MOI = 0.1) at various times (6, 12, 24 h), and then the cells were collected. EV-D68-infected HEK293T cells and RD cells at different MOI (0.01, 0.1, and 1) for 24 h, and then cells were collected. The protein levels of STAT1 and p-STAT1 S727 were measured by western blot. Grayscale analysis of the Western blot results was performed using ImageJ software; three independent grayscale analyses were performed, and the results were averaged to ensure their reliability. qRT-PCR results are presented as the mean and standard deviation from three independent experiments. Statistical significance was analyzed using the Student’s t test (* P < 0.05, ∗ ∗ P < 0.01, ∗ ∗ ∗ P < 0.001, ∗ ∗ ∗ ∗ P < 0.0001, ns, not significant).

Table S1. Primers used in this study
Primer	Sequence (5' - 3')	
q-GAPDH-F	AGATCCCTCCAAAATCAAGTGG	
q-GAPDH-R	GGCAGAGATGATGACCCTTTT	
q-SOCS3-F	CCTGCGCCTCAAGACCTTC	
q-SOCS3-R	GTCACTGCGCTCCAGTAGAA	
q-EVD68-F	GGCAGCCTATCAGGTGGAGAG	
q-EVD68-R	GAGTTTGTATGGCTTCTTCTGGT	
q-ISG15-F	CGCAGATCACCCAGAAGATT	
q-ISG15-R	GCCCTTGTTATTCCTCACCA	
q-RNaseL-F	AGGAGGATCAAGAGCGGCTGA	
q-RNaseL-R	CAAGTGCTTCTTCTCCACTGC	
q- RFX7-F	CAAACCAGCGAACACCAGTG	
q-RFX7-R	TATTGCCTGTTGACGTGCCT