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. 2020 Mar 2;11:1146. doi: 10.1038/s41467-020-14948-z

Fig. 6. Viruses upregulate HOIP and STAT1 linear ubiquitination.

Fig. 6

a RT-qPCR analysis of HOIP mRNA in HT1080 cells challenged with SeV (MOI = 1.0) for the indicated times. b RT-qPCR analysis of HOIP mRNA in HT1080 cells infected with SeV (MOI = 1.0) and then treated with a NF-κB inhibitor PDTC (10 µM) as indicated. c RT-qPCR analysis of HOIP mRNA in HT1080 cells stimulated with IFNα (1000 IU/ml) as indicated. d Western blot analysis of HOIP proteins in HEK293T cells infected with VSV (MOI = 0.1) as indicated. e Immunoprecipitation analysis of the interaction between HOIP and STAT1 in 2fTGH cells infected with VSV (MOI = 0.1) as indicated. f Immunoprecipitation analysis of linear ubiquitination of STAT1 in 2fTGH cells infected with VSV (MOI = 0.1 and 0.5) for 9 h. g Immunoprecipitation analysis of linear ubiquitination of STAT1 in HT1080 cells infected with SeV (MOI = 1.0) as indicated. h Immunoprecipitation analysis of linear ubiquitination of STAT1 in HOIP-WT or HOIP-KO HEK293T cells infected with or without VSV (MOI = 0.1) for 9 h. i Immunoprecipitation analysis of linear ubiquitination of STAT1 in HEK293T cells transfected with Myc-STAT1 (WT or DM) and then infected with VSV (MOI = 0.1) as indicated. j Immunoprecipitation analysis of linear ubiquitination of STAT1 and RT-qPCR analysis of Ifnβ mRNA in 2fTGH cells infected with SeV for the indicated times. k Western blot analysis of pY-STAT1 in HOIP-WT or HOIP-KO HEK293T cells infected with VSV (MOI = 0.1) for 12 h. l RT-qPCR analysis of Ifit1 mRNA in U3A cells transfected with Myc-STAT1 (WT or DM) and then infected with SeV (MOI = 1.0) as indicated. N.S., not significant (p > 0.05) and *p < 0.05, **p < 0.01, ***p < 0.001 (two-tailed unpaired Student’s t test). Data are shown as mean and s.d. of three biological replicates (a–c, l), or are representative of three independent experiments (d–k).