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. 2018 Apr 17;9(29):20386–20398. doi: 10.18632/oncotarget.24910

Figure 5. MCT4 silencing had no effect on metabolic parameters as well as viability, proliferation and death of JEG-3 cells.

Figure 5

(A) Western blot results for MCT4 expression after SLC16A3 knockdown, using 10 nM siRNA, for 72 hours, showing an effective silencing of MCT4 expression. Scramble siRNA was used as negative control and β-actin was used as loading control. Extracellular amounts of glucose (B) and lactate (C) levels were quantified in culture medium obtained after 24 hours, normalized by total biomass (obtained using SRB assay) and are represented as the mean of three independent experiments in triplicate ± SEM. Cell viability was evaluated using SRB assay (D) while cell proliferation was evaluated using BrdU incorporation (E). Results were obtained after 24 hours of cell culture, normalized as the percentage of the negative control (scramble siRNA), and are represented as the mean of three independent experiments in triplicate ± SEM. Cell death was evaluated using flow cytometry analysis after Annexin V/PI staining (F). Results were obtained after 24 hours and are represented by the mean percentage of double stained cells (late apoptosis/necrosis) from two independent experiments ± SEM.