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. 2020 Mar 30;9(4):834. doi: 10.3390/cells9040834

Figure 3.

Figure 3

Putative HBV restriction factors are processed through a CRL complex: (a) HEK293T cells were cultured for 24 h in the presence of increasing concentrations of the CRL inhibitor MLN4924. Protein levels for putative restriction factors PSME4, Zinc Finger E-Box Binding Homeobox 2 (ZEB2), STIM1, and PSME1 and for SMC6 [13,14] in the absence and presence of MLN4924 were determined by western blot assays and normalized either to total protein (indicated by asterisks) or to actin (described in the Materials and Methods section). (b) Graphical representation of western blot shown in panel (a): The level of normalized restriction factor in untreated cells was set to 1.0 and the levels of the same protein in MLN4924-treated cells compared to that. PSME4, ZEB2, and SMC6 showed increasing levels with increasing amounts of MLN4924, indicating they are normally processed through a CRL. STIM1 levels increased with the highest dose (10 μg), while PSME1 levels were not impacted by drug treatment. Results shown are representative of n = 3 independent experiments.