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. 2015 Jun 12;290(31):19044–19054. doi: 10.1074/jbc.M115.637660

FIGURE 4.

FIGURE 4.

Siah2 is required for GPS2 degradation. A, transient transfection of equal amounts of HA-Siah1 and HA-Siah2 in 293T cells prevents the expression of HA-GPS2, whereas coexpressing GPS2 together with TBL1 increases its stabilization. Blotting for tubulin provided a loading control. B, the endogenous level of nuclear GPS2 increases upon Siah1 and Siah2 down-regulation via specific siRNA. Blotting for HDAC2 provided a loading control. C, Western blot analysis of CEs and NEs showing that overexpression of Siah1/2 in 293T cells fails to decrease the expression of a mutant GPS2 lacking the three C-terminal lysines (K254,300,327A). Blotting for tubulin and HDAC2 provided controls for CEs and Nes, respectively. D, Siah2 genetic ablation, as verified by blotting for Siah2, promotes an increased GPS2 protein level in extracts from mouse adipose tissue. Blotting for tubulin provided a loading control.