Fig. 2

MCT4 promotes astrocyte glycolysis and proliferation. (A) Analysis of glucose consumption and lactate production levels in MCT4-knockdown astrocytes (shMCT4) and control astrocytes (shCtl). (B) Primary astrocytes were treated with MCT4 inhibitor CHCA for 24 h, followed by stimulation with 50 ng/mL TNF-α and IL-1β for 24 h. Glucose consumption and lactate production levels were measured. (C) Analysis of lactate production levels in MCT4-overexpressed astrocytes (LV-MCT4), control astrocytes (LV-NC), and non-treated astrocytes (mock). (D) qRT-PCR analysis of MCT4, GLUT1, HK2, Cyclin D1 and Cyclin E in MCT4-knockdown astrocytes and control astrocytes. (E) Effect of MCT4 inhibitor CHCA (1 mM) on the mRNA expression of MCT4, GLUT1, HK2, PKM2 and Cyclin D1 in TNF-α and IL-1β-stimulated astrocytes. (F) Effect of MCT4 inhibitor CHCA on the proliferation of TNF-α and IL-1β stimulated astrocytes was measured by EdU assay. For quantification of EdU, 6 fields were included for each group. (G) Cell proliferation was measured in MCT4 knockdown and control astrocytes. EdU⁺ cells were quantified. (H) Cell proliferation was measured in mock, control-lentivirus and MCT4-overexpressed astrocytes by EdU incorporation assay. EdU⁺ cells were quantified by Image-Pro Plus. Scale bar: 100 μm. Data are represented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 using one-way ANOVA with Dunnett’s multiple comparison test