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. 2019 Sep 18;10:2147. doi: 10.3389/fimmu.2019.02147

Figure 2.

Figure 2

Txnip ubiquitination initiates proteasomal degradation in GAS-infected macrophages. (A) RAW264.7 cells were pretreated with or without MG132 and LAC for 1 h followed by GAS infection, and the expression of Txnip was determined. (B) In the presence or absence of BafA1 and CQ, Txnip expression in GAS-infected cells was detected. GAPDH is used as an internal control. The expression ratios of Txnip to internal controls are shown. (C) In the presence or absence of MG132 (10 μM), RAW264.7 cells were infected with GAS (MOI 10), and Txnip proteins were then immunoprecipitated (IP) followed by subsequent immunoblotting (IB) with anti-Ub and anti-Txnip antibodies. Cell lysate is used as the positive control and the Txnip-Ub polyubiquitination is labeled. Protein molecular weights (MW) are indicated in kilodaltons. Western blot results represent at least two independent experiments. (D) In the presence or absence of MG132 (10 μM), RAW264.7 cells were infected with GAS (MOI 10) followed by the measurement of Trx-1 activity at the indicated hpi. Data are shown as the means ± SD of triplicate cultures. *p < 0.05; **p < 0.01.