The effect of MCU inhibition on the phosphorylation of MAPK and NF-κB pathway.

<p>HepG2 cells were treated with HG for 24 h. After treatment, cells were lysed and protein extracts were immunoblotted with specific antibodies. The effect of MCU inhibition on JNK (A), P38 (B), ERK (C) and IKKα-β (D) phosphorylation has been demonstrated. Data are shown as a mean ± SEM of at least three separate experiment. MCU knockdown cells, SC: Scramble, NG: normal glucose, HG: high glucose, # <0.05, * <0.01, * * <0.001.</p

The effect of high glucose on insulin resistance and mitochondrial calcium homeostasis.

<p>A: The effect of 33mM glucose (HG) on Akt phosphorylation was performed by western blotting method. B&C: The effect of HG on mitochondrial calcium was assessed using the adenoviruses expressing the low-affinity mitochondrial probe. D&G: The effect of HG on cytosolic calcium was evaluated by adenoviruses expressing the cytosolic probe. Data are shown as a mean ± SEM of at least three separate experiment. NG: normal glucose, HG: high glucose, # <0.05, * <0.01, NS = not significance.</p

The effect of MCU inhibition on ROS production in HepG2 cells.

<p>HepG2 cells were treated with HG for 24 h. A: H₂O₂ levels were measured using flow cytometry with DCFH-DA. B: Mitochondrial ROS level using MitoSOX red dye. Data are shown as a mean ± SEM of at least three separate experiment.: MCU knockdown cells, SC: Scramble, NG: normal glucose, HG: high glucose, # <0.05, * <0.01, * * <0.001.</p

Importance of MCU inhibition in high glucose-induced pro-inflammatory and coagulative responses.

<p>HepG2 stable cells were generated by infecting the cells with the supernatants of lentiviruses expressing MCU shRNA. Real-time PCR and western blotting were used to detect MCU mRNA and protein levels in HepG2 stable cells, respectively. A: Protein levels of MCU, B: mRNA level of MCU, C) mitochondrial calcium concentration in MCU-KD cells. D: the effect of MCU inhibition on the mRNA expression of TNF-α (E), IL-6 (F), PAI-1 (G), FGA (H), and FGB(I) were measured using real time PCR. Data are shown as a mean ± SEM of at least three separate experiment. MCU-KD: MCU knockdown cells, SC: Scramble, NG: normal glucose, HG: high glucose, # <0.05, * <0.01, * * <0.001.</p