Fig. 3

β-Hydroxybutyric acid-induced mitochondrial dysfunction in fGCs. A Representative images of the mitochondrial (red arrows) morphology of fGCs after BHBA treatment by TEM (scale bars = 0.5 μm). Normal mitochondria are marked with blue arrows, abnormal mitochondria are marked with red arrows, and autolysosomes are marked with stars. B The abnormal rate of mitochondria was statistically analyzed in five images for each treatment group (n = 5). C–D Analysis of MPTP detection and fluorescence intensity after BHBA treatment (scale bars = 20 μm, n = 8). Calcein AM showed strong green fluorescence after incubation with cells. After further incubation with CoCl₂ (Calcein AM + CoCl₂), the green fluorescence of calcein in the cytoplasm was quenched, and only mitochondria showed green fluorescence, with Ionomycin as a negative control (Calcein AM + CoCl₂). E–F Representative images and fluorescence intensity analysis showing intracellular Ca²⁺ levels after BHBA treatment (scale bars = 100 μm, n = 5). G–H Representative images and fluorescence intensity analysis of mitochondrial staining under BHBA stimulation (scale bars = 25 μm, n = 8). I Bar graph showing intracellular ATP levels after BHBA stimulation (n = 3). For Fig. 3B, D, F, H and I, we used Student t-tests for significant difference analysis (^*P < 0.05; ^**P < 0.01)