Fig. 3: ASS1 promotes PHGDH degradation by ubiquitination. | Cell Death & Disease

Fig. 3: ASS1 promotes PHGDH degradation by ubiquitination.

From: ASS1 inhibits triple-negative breast cancer by regulating PHGDH stability and de novo serine synthesis

Fig. 3

A, B Immunoblotting assays examining the expression levels of PHGDH in MDA-MB-468 cells with ASS1 knockout (A) or ASS1 overexpression (B) treated with DMSO or 10 μM MG132. Ub was used as a positive control. C, D Immunoblotting assays assessing the expression levels of PHGDH in MDA-MB-468 cells with ASS1 knockout (C) or ASS1 overexpression (D) treated with DMSO or CHX (200 μM). GAPDH was used as a loading control. For each cell, the quantification of the relative endogenous PHGDH protein levels from three independent experiments is shown. Data are means ± SD with p-values based on unpaired T-tests (*p < 0.05). E Immunoprecipitation (IP)-IB of ubiquitinated PHGDH using 293 T cells transfected with ASS1-Myc, PHGDH-Flag, or HA-Ub and treated with MG132 (10 μM) for 8 h before harvest. F Immunoprecipitation (IP)-IB of ubiquitinated PHGDH using MDA-MB-468 Con or ASS1 knockout cells transfected with PHGDH-Flag, or HA-Ub and treated with MG132 (10 μM) for 8 h before harvest.

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