MCT4 Promotes Tumor Malignancy in F98 Glioma Cells

Figure 3

Malignancy in MCT4 overexpression and knockdown F98 glioma cells. (a) Representative images of cell density of con, MCT4, and MCT4^KD F98 cells as well as MCT4 cells treated with 100 μM pCMBS or 150 μM Phl. Scale bar represents 200 μm. (b) Proliferation of con, MCT4, and MCT4^KD F98 cells as well as MCT4 cells treated with 100 μM pCMBS or 150 μM Phl and cultured for 48 h and 72 h as determined by cell counting. Data were normalized to con after 48 h. Statistical analysis was performed by two-way ANOVA with Bonferroni posttest (, mean ± SEM, n = 3). (c) Cell viability of con, MCT4, and MCT4^KD F98 cells as well as MCT4 cells treated with 100 μM pCMBS or 150 μM Phl and cultured for 48 h and 72 h as determined by MTT assay. Data were normalized to con after 48 h Statistical analysis was performed by two-way ANOVA with Bonferroni posttest (, mean ± SEM, n = 3). (d) Actin cytoskeleton morphology in con, MCT4, and MCT4^KD F98 cells (green), visualized by phalloidin staining (red). Arrows indicate cytoskeleton networks between cells. Scale bar represents 20 μm. Quantification of MFI of the red channel is shown below. Statistical analysis was performed by one-way ANOVA with Tukey’s posttest (, mean ± SEM, n ≥ 3). (e) Migration of con, MCT4, and MCT4^KD F98 cells as well as MCT4 cells treated with 100 μM pCMBS or 150 μM Phl after 12 h and 24 h as determined by scratch assay (dashed lines). Scale bar represents 200 μm. Quantification was performed by measuring the scratch length with Fiji and normalizing it to 0 h and con. Statistical analysis was performed by two-way ANOVA with Bonferroni posttest (; , mean ± SEM, n = 3). (f) Colony formation of con, MCT4, and MCT4^KD F98 cells as well as MCT4 cells treated with 100 μM pCMBS or 150 μM Phl after 7 d in 3D cell culture. Images in the upper row depict raw data, whereas images in the row below are postprocessed with Fiji. Scale bar represents 200 μm. Quantification was performed with Fiji and normalized to con. Statistical analysis was performed by one-way ANOVA with Bonferroni posttest (, , and , mean ± SEM, n = 3).

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