Extended Data Fig. 7: Nuclear PHGDH promotes tumorigenesis.
From: The alternative activity of nuclear PHGDH contributes to tumour growth under nutrient stress
In a-h, a total of 1 × 107 SW1990 (a and c) or 5 × 106 LM3 (b, d-h) cells expressing PHGDH sg-RNA (a and b) or c-Jun sg-RNA (h) and indicated Flag-rPHGDH (c-e) or Flag-rc-Jun (f-g) were subcutaneously injected into the athymic nude mice. Tumor volumes were measured by using length (a) and width (b) and calculated using the following equation: V = ab2/2. Data represent the mean ± s.d. (n = 7), statistical analysis was performed using the two-tailed Student’s t-test. (a-e) Representative tumor xenografts (left panels) and quantification of tumor volumes were shown (middle panels); lysates harvested from SW1990 or LM3 cells that were cultured for 6 h in the glucose-free DMEM and tumor tissues were subjected to immunoblotting analyses (right panels). (f-h) NCT503 (40 mg/kg/day) was used to administrate mice 7 days post subcutaneous injection of tumor cells. Representative tumor xenografts (left panels) and quantification of tumor volumes were shown (middle panels); lysates were harvested from LM3 cells that were cultured for 6 h in the glucose-free DMEM and tumor tissues were subjected to immunoprecipitation or immunoblotting analyses (right panels). (i) The PHGDH pS371 (upper panel) and PHGDH pS55 (bottom panel) antibodies specificities were validated using IHC analyses with specific blocking PHGDH pS371 or PHGDH pS55 peptides. Scale bar: 50 μm.
