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. 2019 Aug 20;23(10):6907–6918. doi: 10.1111/jcmm.14574

Figure 5.

Figure 5

Proteasomal degradation of ANXA2 in the knockdown of S100A11. A, Knockdown of S100A11 decreased the expression of ANXA2, while overexpression of S100A11 elevated the expression of ANXA2. B, U87 transfected with shCtrl, sh‐S100A11‐1 or sh‐S100A11‐2, and the cells were treated with or without 10 μM of MG132, and the cell lysates were immunoblotted. C, U87 transfected with shCtrl, or sh‐S100A11‐1 were treated with 50 μg/mL cycloheximide. Whole‐cell lysates were harvested at the indicated times. Knockdown S100A11 decreased the half‐life of ANXA2. D, The interaction between S100A11 and ANXA2 was confirmed by co‐immunoprecipitation in U87 and U251 cells. E, U87 cells were co‐transfected with plasmids expressing HA‐Ub and Myc‐ANXA2 together with either shCtrl or sh‐S100A11‐1. U251 cells were co‐transfected with plasmids expressing HA‐Ub and Myc‐ANXA2 together with either Vector or S100A11 plasmid. Cells were treated with MG132 for 6 h before cell lysates were immunoprecipitated using a denature IP protocol to pull down ANXA2 protein, and the polyubiquitinated ANXA2 protein was detected by anti‐HA antibody