Fig. 4

210M induces glycolysis and lactate export of primary human astrocytes. A miRPathDB was used to predict pathways targeted by miR-210. The pathways with the most hits are listed in descending order, and the p-value of each pathway is included inside the respective bar. B Oxygen Consumption Rate (OCR) was measured in primary human astrocytes 48 h after transfection with 210S or 210M. The basal oxygen consumption (before oligomycin) and maximal respiratory capacity (between FCCP and Rotenone/Antimycin A) are presented in the histogram. n = 3 with 10 technical replicates for OCR experiments. C Extracellular Acidification Rate (ECAR) was measured in primary human astrocytes 48 h after transfection with 210S or 210M. The basal (before glucose), normal glycolysis (between glucose and oligomycin) and glycolytic capacity (between oligomycin and 2-DG addition) are presented in the histogram. n = 4 with 20 technical replicates for ECAR experiments. D RT-qPCR assessment of MCT4 in 210M-transfected cells relative to 210S-transfected cells 48 h after transfection. Mean ± SEM of 8 donors. E RT-qPCR assessment of GPD1L in 210M-transfected cells relative to 210S-transfected cells 48 h after transfection. Mean ± SEM of 8 donors. F Immunoblotted bands of GPD1L and Beta-tubulin (Beta-Tub) proteins and their quantification in 210M-transfected relative to 210S-transfected cells 48 h after transfection. Mean ± SEM of 6 donors. G Primary human astrocytes were transfected with 210S or 210M. After 48 h, cells were washed, and the concentration of lactate was measured in media that was incubated with astrocytes for 2 h. Mean ± SEM of 8 donors. Statistical comparisons were made by two-way ANOVA with Sidak’s correction (B and C) or by a paired t-test (D–H). *p < 0.05; **p < 0.01; ***p < 0.001