adaptor apoptosis  (Bioss)

Journal: Theranostics

Article Title: p66Shc Contributes to Liver Fibrosis through the Regulation of Mitochondrial Reactive Oxygen Species

doi: 10.7150/thno.29620

Figure Lengend Snippet: Primer sequences

Article Snippet: The relevant primary antibodies that were specific for p66Shc (BD Biosciences, USA); cleaved caspase-1 (Affinity Biosciences, USA); Col1a1 (Abcam Ltd., Cambridge, UK);IL-18 (ABclonal Biotechnology, Wuhan, China); α-SMA, NLRP3, adaptor apoptosis- associated speck-like protein (ASC), superoxide dismutase 2 (SOD2), mitochondrial uncoupling protein 1 (UCP1), IL-1β, Cytochrome c , β-actin and voltage-dependent ion channel (VDAC, Proteintech Group, Wuhan, China), followed by secondary antibodies incubation.

Techniques:

Journal: Theranostics

Article Title: p66Shc Contributes to Liver Fibrosis through the Regulation of Mitochondrial Reactive Oxygen Species

doi: 10.7150/thno.29620

Figure Lengend Snippet: p66Shc expression is correlated with liver fibrosis in mice. (A) p66Shc, Col1a1, α-SMA protein levels, n=3. (B) p66Shc, Col1a1, α-SMA mRNA levels, n=6. (C) H&E staining, Masson staining, IHC staining for α-SMA and p66Shc, and dual immunofluorescence staining for p66Shc and α-SMA, as well as for p66Shc and Col1a1. Scale bar, 200 μm. (D) Pearson's correlation analyses of p66Shc IHC scores with Ishak scores of Masson staining (r=0.798, P<0.01). (E) Pearson's correlation analyses of p66Shc mRNA with α-SMA mRNA (r=0.696, P<0.05). ## P<0.01.

Article Snippet: The relevant primary antibodies that were specific for p66Shc (BD Biosciences, USA); cleaved caspase-1 (Affinity Biosciences, USA); Col1a1 (Abcam Ltd., Cambridge, UK);IL-18 (ABclonal Biotechnology, Wuhan, China); α-SMA, NLRP3, adaptor apoptosis- associated speck-like protein (ASC), superoxide dismutase 2 (SOD2), mitochondrial uncoupling protein 1 (UCP1), IL-1β, Cytochrome c , β-actin and voltage-dependent ion channel (VDAC, Proteintech Group, Wuhan, China), followed by secondary antibodies incubation.

Techniques: Expressing, Staining, Immunohistochemistry, Immunofluorescence

Journal: Theranostics

Article Title: p66Shc Contributes to Liver Fibrosis through the Regulation of Mitochondrial Reactive Oxygen Species

doi: 10.7150/thno.29620

Figure Lengend Snippet: p66Shc silencing attenuates liver fibrosis in mice. p66Shc silencing was induced via lentivirus delivered to C57BL/6 mice exposed to CCl 4 (2 ml/kg). (A) Liver p66Shc mRNA expression, n=6. (B) Liver p66Shc, SOD2, UCP1, Col1a1, α-SMA protein, n=3. (C) H 2 O 2 content, n=8. (D) SOD activity, n=8. (E) Cytochrome c expression in the cytoplasm and mitochondria, n=3. (F) H&E and Masson staining. Scale bar, 200 μm. (G) Ishak score of Masson staining. (H) Serum ALT and AST levels, n=8. (I) Liver NLRP3 inflammasome protein expression, n=3. (J) Liver CTGF and TIMP1 mRNA levels, n=6. ## P<0.01, # P<0.05.

Article Snippet: The relevant primary antibodies that were specific for p66Shc (BD Biosciences, USA); cleaved caspase-1 (Affinity Biosciences, USA); Col1a1 (Abcam Ltd., Cambridge, UK);IL-18 (ABclonal Biotechnology, Wuhan, China); α-SMA, NLRP3, adaptor apoptosis- associated speck-like protein (ASC), superoxide dismutase 2 (SOD2), mitochondrial uncoupling protein 1 (UCP1), IL-1β, Cytochrome c , β-actin and voltage-dependent ion channel (VDAC, Proteintech Group, Wuhan, China), followed by secondary antibodies incubation.

Techniques: Expressing, Activity Assay, Staining

Journal: Theranostics

Article Title: p66Shc Contributes to Liver Fibrosis through the Regulation of Mitochondrial Reactive Oxygen Species

doi: 10.7150/thno.29620

Figure Lengend Snippet: p66Shc contributes to HSC activation in vitro . Primary HSCs were transfected with p66Shc siRNA (A-C) or pcDNA-p66Shc (D, E) and then exposed to TGF-β1. (A) p66Shc, Col1a1, α-SMA protein, n=3. (B) CTGF and TIMP1 mRNA, n=6. (C) Dual immunofluorescence of p66Shc and α-SMA. Representative immunofluorescence images in the left panel. Scale bar, 200 μm. Representative cell area in the right panel. (D) p66Shc, Col1a1, α-SMA protein expression, n=3. (E) CTGF and TIMP1 mRNA expression, n=6. ## P<0.01.

Article Snippet: The relevant primary antibodies that were specific for p66Shc (BD Biosciences, USA); cleaved caspase-1 (Affinity Biosciences, USA); Col1a1 (Abcam Ltd., Cambridge, UK);IL-18 (ABclonal Biotechnology, Wuhan, China); α-SMA, NLRP3, adaptor apoptosis- associated speck-like protein (ASC), superoxide dismutase 2 (SOD2), mitochondrial uncoupling protein 1 (UCP1), IL-1β, Cytochrome c , β-actin and voltage-dependent ion channel (VDAC, Proteintech Group, Wuhan, China), followed by secondary antibodies incubation.

Techniques: Activation Assay, In Vitro, Transfection, Immunofluorescence, Expressing

Journal: Theranostics

Article Title: p66Shc Contributes to Liver Fibrosis through the Regulation of Mitochondrial Reactive Oxygen Species

doi: 10.7150/thno.29620

Figure Lengend Snippet: p66Shc triggers NLRP3 inflammasome activation through regulation of mitochondrial ROS production. (A) Primary HSCs were stimulated with MSU and mitochondrial complex inhibitor (rotenone, TTFA and antimycin A). NLRP3 and IL-1β protein expression, n=3. (B) p66Shc overexpression was carried out in primary HSCs by pcDNA-p66Shc vector and then rotenone or antimycin A was stimulated. NLRP3 and IL-1β protein expression, n=3. (C-E) p66Shc overexpression was applied in primary HSCs by pcDNA-p66Shc vector transfection and then mito-TEMPO was stimulated under TGF-β1 treatment. (C) Representative fluorescence images of MitoSOX. (D) NLRP3, IL-1β, Col1a1, α-SMA protein levels, n=3. (E) CTGF and TIMP1 mRNA expression, n=6. (F-G) Primary HSCs were co-transfected with pcDNA-p66Shc and NLRP3 siRNA following TGF-β1 challenge. (F) NLRP3, IL-1β, Col1a1, α-SMA protein levels, n=3. (G) CTGF and TIMP1 mRNA expression, n=6. ## P<0.01, # P<0.05.

Article Snippet: The relevant primary antibodies that were specific for p66Shc (BD Biosciences, USA); cleaved caspase-1 (Affinity Biosciences, USA); Col1a1 (Abcam Ltd., Cambridge, UK);IL-18 (ABclonal Biotechnology, Wuhan, China); α-SMA, NLRP3, adaptor apoptosis- associated speck-like protein (ASC), superoxide dismutase 2 (SOD2), mitochondrial uncoupling protein 1 (UCP1), IL-1β, Cytochrome c , β-actin and voltage-dependent ion channel (VDAC, Proteintech Group, Wuhan, China), followed by secondary antibodies incubation.

Techniques: Activation Assay, Expressing, Over Expression, Plasmid Preparation, Transfection, Fluorescence

Journal: Theranostics

Article Title: p66Shc Contributes to Liver Fibrosis through the Regulation of Mitochondrial Reactive Oxygen Species

doi: 10.7150/thno.29620

Figure Lengend Snippet: p66Shc upregulation and NLRP3 inflammasome activation are involved in human liver fibrosis. Relative protein expression and mRNA were measured in patients with liver fibrosis. (A) p66Shc mRNA levels. (B) p66Shc, SOD2, UCP1, Col1a1, α-SMA protein levels. (C) NLRP3 inflammasome protein levels. (D) CTGF and TIMP1 mRNA levels. n=12. ## P<0.01, # P<0.05.

Article Snippet: The relevant primary antibodies that were specific for p66Shc (BD Biosciences, USA); cleaved caspase-1 (Affinity Biosciences, USA); Col1a1 (Abcam Ltd., Cambridge, UK);IL-18 (ABclonal Biotechnology, Wuhan, China); α-SMA, NLRP3, adaptor apoptosis- associated speck-like protein (ASC), superoxide dismutase 2 (SOD2), mitochondrial uncoupling protein 1 (UCP1), IL-1β, Cytochrome c , β-actin and voltage-dependent ion channel (VDAC, Proteintech Group, Wuhan, China), followed by secondary antibodies incubation.

Techniques: Activation Assay, Expressing