Fig 1: Morphological changes of mitochondria caused by high glucose. The white arrows indicate the mitochondria of HTR-8/SVneo cells. The mitochondria broke into nodules or fragments in the HG group, lacking continuity; those in the NG group were intact and clear. NG, HTR-8/SVneo cells cultured in 5.6 mM glucose; HG, cells cultured in 25 mM glucose. Green, TXNIP-positive cells; blue, DAPI for nuclear staining. Scale bar 25 µm. Fixed tissue images were captured in a Leica confocal microscope at 40×
Fig 2: Model of the cell apoptosis procedure that happens in the HTR-8/SVneo cells with high expression of TXNIP treated with HG. TXNIP expression is strongly induced in HTR-8/SVneo cells treated with HG. TXNIP is primarily located in the cytoplasm and nucleus. TXNIP translocates into mitochondria where it interacts with TXN when cellular stress occurs. The combination of TXNIP with TXN attenuates the reducing ability of TXN, increases ROS accumulation, and results in mitochondrial dysfunction (morphological change and inhibition of ATP generation). TXNIP can also release ASK1 from the TXN–ASK1 complex. The released ASK1 turns on the pathway of apoptosis, which leads to cleavage of caspase-3 and cell apoptosis
Fig 3: Effects of MitoQ on NLRP3, TRX and TXNIP expression and intracellular ROS production in HK-2 cells exposed to different concentrations of D-glucose. A: Representations of intracellular ROS levels (A, upper panel), MMP (A, middle panel) and mitochondrial ROS levels (A, bottom panel) in HK-2 cells subjected to HG treatment with or without MitoQ pretreatment. B-F: Quantification of intracellular ROS production as measured with DCFDA staining (B), MMP as measured with TMRE staining (C), mitochondrial ROS production as measured with MitoSox Red staining (D), cellular H2O2 production (E) and Ca2+ loading for MPTP opening (F) in HK-2 cells treated with or without different concentrations of MitoQ. G-J: WB and densitometric analysis of NLRP3, TRX, TXNIP, Caspase-1, IL-1β, Collagen I and FN expression in HK-2 cells exposed to different concentrations of D-glucose and treated with or without MitoQ. Values are the mean ± SE, *P < 0.05, vs control group; #P < 0.05 vs HG group. n = 3.
Fig 4: Overexpression of TXNIP induces mitochondrial dysfunction in HTR-8/SVneo cells. HTR-8/SVneo cells were transfected with pCMV3-TXNIP or pCMV3-NCV (0.28 μg/mL). Mitochondrial morphology was assessed at 6 h after TXNIP was transfected. a Overexpression of TXNIP changed the mitochondria of the cells morphologically. The white arrows indicate mitochondria of HTR-8/SVneo cells. The mitochondrial fragmentation observed in the OE-TXNIP group was higher than that in the NC and the control group. Green, cells expressing TXNIP; red, Mito Tracker; blue, DAPI for nuclear staining. Scale bar 25 µm. Fixed cell images were captured in a Leica confocal microscope at ×40. b Mitochondrial membrane potential decreased in the OE-TXNIP group. The mitochondrial membrane potential marked by 200 mM JC-1. Overexpression of TXNIP decreased the mitochondrial membrane potential to the lowest point at 3 h after TXNIP was transfected. Red fluorescence represents cells with intact mitochondrial membranes, and green fluorescence represents the cells with loss of mitochondrial membrane potential. Images were captured in Biotek at ×20. c The ratio of red fluorescence intensity to green fluorescence intensity was analyzed to quantify the mitochondrial membrane potential (ΔΨ) of the cells. d Analysis of the correlation between TXNIP expression and mitochondrial membrane potential (ΔΨ). The experiments were replicated independently at least three times. Results were expressed as mean ± SEM. ###p < 0.001 (Tukey’s test) vs NC group (transfected with pCMV3-NCV). ***p < 0.001 vs control group (without transfection)
Fig 5: Protein expression of TXNIP in the placenta syncytioctrophoblasts of women with GDM was higher than that of the normal group. Frozen placenta sections from the GDM group or normal group were stained with TXNIP antibody. The protein expression of TXNIP in villous tissue of placentas from both groups were investigated via confocal photomicrographs. Green, TXNIP-positive cells; blue, DAPI for nuclear staining. Scale bar 75 µm. Fixed tissue images were captured in a Leica confocal microscope at 40×. The white arrows indicate syncytioctrophoblasts, and the fluorescence intensity in the GDM group was higher than that in the normal group, ***p < 0.001 (Tukey’s test) vs normal group
Supplier Page from Abcam for Anti-TXNIP antibody [EPR14774] - BSA and Azide free