Fig. 6.

STAT deficiency induces a glycolytic metabolism in TCCs. (A, 1) shCT and shSTAT3 TCCs (8505C and TPC-1) were grown for 96 and 48 h, respectively, with RPMI (1% serum) and different concentrations of the hypoxia-mimetic CoCl₂. Growth was determined by the sulpharhodamine B assay. (A, 2) shCT and shSTAT3 cell lines (8505C and TPC-1) were treated with CoCl₂ (100 μM) for 24 h. Total protein was probed with the indicated antibodies. (B and C) Cells were grown in plain RPMI for 48 h with or without CoCl₂ (100 μM). (B) Glucose uptake was determined by subtracting the glucose concentration in the supernatant from the initial concentration in fresh RPMI. Lactate was measured in the supernatant of the cell. Results were normalized to protein amount (milligrams). Bars represent mean ± SEM (n = 3). (C) The oxygen consumption rate and mitochondrial membrane potential (JC-1) were measured in equal numbers of 8505C shCT and shSTAT3 cells (mean ± SEM; n = 3). (D) 8505C shCT and shSTAT3 cell lines were treated with CoCl₂ (100 μM) for 24 h, and the RNA levels of the indicated glycolytic enzymes (PDK1 and GLUT1/3) were determined and normalized to β-actin. Bars are mean ± SEM (n = 3). *P < 0.05, **P < 0.01, ***P < 0.0001.