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. 2017 Nov 20;13(11):e1006693. doi: 10.1371/journal.ppat.1006693

Fig 8. iRhom2 inhibits virus-triggered MAD of VISA.

Fig 8

(A) Co-immunoprecipitation and immunoblot analysis using the indicated antibodies with lysates of HEK293T cells transfected with the indicated plasmids. (B) Co-immunoprecipitation and immunoblot analysis using the indicated antibodies with lysates of HEK293T cells infected with SeV for the indicated times. (C) Detection of dose-dependent effects of iRhom2 on the level of MARCH5 in HEK293T cells transfected with the indicated plasmids by immunoblot analysis with the indicated antibodies. (D) The experiments were similarly performed as in (C) except that cells were untreated or treated with MG132 (100 μM) for 6 h before immunoblot analysis. +: 0.5μg, ++: 1μg. (E) Immunoblot analysis of the indicated proteins in iRhom2+/+ and iRhom2–/–MEFs infected with SeV for the indicated times. (F) qPCR analysis of MARCH5 mRNA in HEK293T cells transfected with iRhom2-shRNA, selected with puromycin (1 μg/ml) for 36 h and then infected with SeV for the indicated times. (G) Immunoblot analysis of endogenous VISA in wild-type and RNF5-KO HEK293T cells transfected with the indicated shRNA plasmids for 2 days and then infected with SeV for the indicated times. (H) qPCR analysis of IFNB1 and IP10 mRNAs in HEK293T cells transfected with the indicated shRNA plasmids, selected with puromycin (1 μg/ml) for 2 days and then infected with SeV for 5 h. (I) Immunoblot analysis of the indicated proteins in VISA-KO HEK293T cells transfected with the indicated plasmids for 24 h. *P < 0.05; **P < 0.01 (unpaired t-test). Data are representative of three experiments with similar results.