Journal: International Journal of Molecular Sciences

Article Title: Heat Shock Protein 60 Restricts Release of Mitochondrial dsRNA to Suppress Hepatic Inflammation and Ameliorate Non-Alcoholic Fatty Liver Disease in Mice

doi: 10.3390/ijms23010577

Figure Lengend Snippet: Analyses of the protein of TLR3 ( A ), MDA5 ( B ), and p-IRF3 ( C ), the mRNA of the il-6 ( D ) and il1-il1β ( E ) expressions, and the ( F ) mitochondria DNA copy number per cell in the WT and HSP60Tg mice livers following HFD or CD for 26 weeks. Data calculated from five to eight samples per group are expressed as the mean ± SE. * p < 0.05, ** p < 0.01 and *** p < 0.001 between the indicated groups. WT, wild-type mice; HFD, high-fat diet; HSP60Tg, mice harboring overexpression of heat shock protein 60; CD, chow diet; TLR3, toll-like receptor 3; MDA5, melanoma differentiation-associated gene 5; p-IRF3, phosphorylated-interferon regulatory factor 3; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Following electrophoresis, it was transferred to a PVDF membrane (Millipore, MA), and the blots were incubated with primary antibodies at 4 °C overnight against HSP60 (sc-1052, Santa Cruz, CA, USA), TLR3 (ab62566, abcam, Cambridge, UK), MDA5 (#5321, Cell Signaling, MA, USA), p-IRF3 (orb6225, Biorbyt, Cambridge, UK), IRF3 (ab68481, abcam, Cambridge, UK) and GAPDH (60004-1-lg, PROTEINTECH, IL, USA) for protein control.

Techniques: Over Expression

Journal: International Journal of Molecular Sciences

Article Title: Heat Shock Protein 60 Restricts Release of Mitochondrial dsRNA to Suppress Hepatic Inflammation and Ameliorate Non-Alcoholic Fatty Liver Disease in Mice

doi: 10.3390/ijms23010577

Figure Lengend Snippet: Analyses of HSP60 ( A ), TLR3 ( B ), MDA5 ( C ), and p-IRF3 ( D ) on the 10th culture day of HepG2 after treatment with a HSP60 overexpression vector for 24 h. Data are expressed as the mean ± SE of the four to six samples per group. ** p < 0.01 and *** p < 0.001 between the indicated groups. TLR3, toll-like receptor 3; MDA5, melanoma differentiation-associated gene 5; P-IRF3, phosphorylated-interferon regulatory factor 3; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Following electrophoresis, it was transferred to a PVDF membrane (Millipore, MA), and the blots were incubated with primary antibodies at 4 °C overnight against HSP60 (sc-1052, Santa Cruz, CA, USA), TLR3 (ab62566, abcam, Cambridge, UK), MDA5 (#5321, Cell Signaling, MA, USA), p-IRF3 (orb6225, Biorbyt, Cambridge, UK), IRF3 (ab68481, abcam, Cambridge, UK) and GAPDH (60004-1-lg, PROTEINTECH, IL, USA) for protein control.

Techniques: Over Expression, Plasmid Preparation

Journal: International Journal of Molecular Sciences

Article Title: Heat Shock Protein 60 Restricts Release of Mitochondrial dsRNA to Suppress Hepatic Inflammation and Ameliorate Non-Alcoholic Fatty Liver Disease in Mice

doi: 10.3390/ijms23010577

Figure Lengend Snippet: Proposed model of heat shock protein 60 restricts release of mitochondrial dsRNA to suppress hepatic inflammation and ameliorate non-alcoholic steatohepatitis in mice. High-fat diet (HFD) composed of 60% calories from fat can induce an obesity animal model that is characterized by steatohepatitis, glucose intolerance, and insulin resistance. Gain of heat shock protein 60 (HSP60) alleviated not only HFD-induced body weight gain, fat accumulation, and hepatocellular steatosis, but also glucose tolerance and insulin resistance. Furthermore, overexpression of HSP60 in the HFD group resulted in inhibited release of mitochondrial dsRNA (mt-dsRNA) compared to wild-type mice. In addition, overexpression of HSP60 also inhibited the activation of toll-like receptor 3 (TLR3), melanoma differentiation-associated gene 5 (MDA5), and phosphorylated-interferon regulatory factor 3 (p-IRF3), as well as inflammatory biomarkers such as mRNA of il-1β and il-6 expression in the liver in response to HFD.

Article Snippet: Following electrophoresis, it was transferred to a PVDF membrane (Millipore, MA), and the blots were incubated with primary antibodies at 4 °C overnight against HSP60 (sc-1052, Santa Cruz, CA, USA), TLR3 (ab62566, abcam, Cambridge, UK), MDA5 (#5321, Cell Signaling, MA, USA), p-IRF3 (orb6225, Biorbyt, Cambridge, UK), IRF3 (ab68481, abcam, Cambridge, UK) and GAPDH (60004-1-lg, PROTEINTECH, IL, USA) for protein control.

Techniques: Animal Model, Over Expression, Activation Assay, Expressing