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. 2021 Feb 22;163(3):278–292. doi: 10.1111/imm.13315

Figure 4.

Figure 4

MID1 regulates protein stability of IRF3. (a) RT‐qPCR analysis of endogenous IRF3 mRNA in HEK293T cells transfected with shCON or two shMID1. (b) RT‐qPCR analysis of endogenous IRF3 mRNA in HEK293T cells transfected with increased amounts of FH‐MID1. Flag‐MID1 protein levels were detected by immunoblotting using an anti‐Flag antibody. (c) HEK293T cells were transfected with HA‐IRF3, together with or without FH‐MID1. Then, cells were treated with CHX (50 mg/ml) for different times. Whole‐cell extracts were analysed by immunoblotting using the indicated Abs. (d) HEK293T cells were transfected with HA‐IRF3, together with or without FH‐MID1. Then, cells were treated with MG132 (10 mm) for 12 h. Whole‐cell extracts were analysed by immunoblotting using the indicated Abs (left). HEK293T cells transfected with HA‐IRF3 and (or) Myc‐MID1 were treated with methylamine hydrochloride (MA; 20 mm) for 12 h. Whole‐cell lysates were analysed by immunoblotting using the indicated Abs (right).