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. 2021 Feb 4;12(12):933–945. doi: 10.1093/jmcb/mjaa043

Figure 3.

Figure 3

HDAC4 positively regulates the level of IκBα protein. (A) Effect of HDAC4 on the level of IκBα in HEK293T cells. Cells (4 × 105) were transfected with the indicated plasmids for 36 h and then were treated with TNFα (20 ng/ml) for the indicated time or left untreated. The cells were lysed, and the lysates were analyzed by immunoblotting with the indicated antibodies. (B) Effect of HDAC4 knockdown on the level of IκBα proteins. The control or HDAC4-knockdown cells (1 × 105) were treated with TNFα (20 ng/ml) for the indicated time or left untreated. (C) Effect of HDAC4 on the stability of IκBα. The HDAC4-knockdown cells (4 × 105) were transfected with the indicated plasmids or treated with the indicated inhibitors MG132 (1 µg/ml), 3-MA (60 µM), and CQ (50 µM) for 4 h before immunoblotting analysis was performed. (D) HDAC4 enhances the interaction between IκBα and NF-κB. HEK293T cells (2 × 106) were transfected with the indicated plasmids (2 µg each). Cell lysates were incubated with a Flag antibody. Bound proteins were analyzed by immunoblotting with anti-HA, anti-Flag, and anti-HDAC4. Data are representative of three independent experiments with similar results.