Journal: PLOS Biology

Article Title: Optineurin binding to the novel interacting partner Junction plakoglobin prevents muscle atrophy in mice

doi: 10.1371/journal.pbio.3003581

Figure Lengend Snippet: RNA sequencing was performed on TA muscle of Optn KD mice and controls. (A) Volcano plot of significantly up (red) and downregulated genes (blue). Not significantly changed genes were indicated in gray. The representative genes related PI3K-AKT signaling pathway and catabolic metabolism signaling pathway were labeled on the volcano plot. Red and blue highlighted fold changes of 1.2 and −1.2. P value < 0.05. (B) KEGG pathway enrichment analysis of significantly changed genes in (A). (C) Heatmap showing expression changes in PI3K-AKT and catabolic metabolism signaling pathway-related genes in TA muscle from Optn KD and control mice by RNA-seq. (D, E) Representative immunoblotting analysis (D) and quantification (E) of PI3K-AKT pathway in scramble shRNA or sh Optn TA muscle ( n = 3 mice in each group). Data are presented as mean ± standard error of the mean (SEM). * P < 0.05 vs. control. The underlying data for this figure can be found in . The Original blot for this figure can be found in .

Article Snippet: To verify the activation of PI3K-AKT signaling pathway in vivo, the TA muscle of scrambled shRNA or sh Optn was injected with 20 ul the specific PI3K activator 740-YP (30 μM, HY-P0175, MedChem Express, Shianghai, PRC) or DMSO per mouse for 4 weeks.

Techniques: RNA Sequencing, Labeling, Expressing, Control, Western Blot, shRNA

Journal: PLOS Biology

Article Title: Optineurin binding to the novel interacting partner Junction plakoglobin prevents muscle atrophy in mice

doi: 10.1371/journal.pbio.3003581

Figure Lengend Snippet: (A) Schematics for pharmacological activation of PI3K-AKT signaling pathway by 740-YP in Optn -KD mice. Four weeks after intramuscular injection of AAV scramble shRNA or AAV-sh Optn to TA muscle, mice were then treated with 30 μM 740-YP per day for 4 weeks. (B, C) Physical performance was evaluated in mice by a treadmill exhaustion test ( n = 5 mice in each group). Two parameters were measured with this test: (B) Time (Left panel) and Running distance (Right panel) to exhaustion (Survival plot showing the percentage of mice running at indicated time points and distances). (C) Quantification of mean duration (left panel) and distance of run to exhaustion (Right panel) ( n = 5 mice in each group). (D) Comparison of representative samples of dissected TA muscle in control or Optn -KD mice with 740-YP treatment. (E) Quantification of TA muscle mass in (D) ( n = 5 mice in each group). (F) Representative H&E and laminin staining of TA muscle in control or Optn KD mice with 740-YP treatment ( n = 5 mice in each group). Scale bar: 100 μm. (G) Representative immunoblotting analysis of muscle atrophy markers (Atrogin-1 and Murf-1) and PI3K-AKT pathway in TA muscle from control or Optn KD mice with 740-YP treatment ( n = 3 mice in each group). Data are presented as mean ± standard error of the mean (SEM). * P < 0.05 vs. control. The underlying data for this figure can be found in . The Original blot for this figure can be found in .

Article Snippet: To verify the activation of PI3K-AKT signaling pathway in vivo, the TA muscle of scrambled shRNA or sh Optn was injected with 20 ul the specific PI3K activator 740-YP (30 μM, HY-P0175, MedChem Express, Shianghai, PRC) or DMSO per mouse for 4 weeks.

Techniques: Activation Assay, Injection, shRNA, Comparison, Control, Staining, Western Blot

Journal: PLOS Biology

Article Title: Optineurin binding to the novel interacting partner Junction plakoglobin prevents muscle atrophy in mice

doi: 10.1371/journal.pbio.3003581

Figure Lengend Snippet: (A, B) Immunoprecipitation analysis of JUP and PI3 Kinase p85 in Optn -overexpressing (A) or KD (B) C2C12 cells. The immunoprecipitation analysis was performed in Optn -overexpressing or KD C2C12 cells at 4 d post-differentiation, incubated with anti-JUP antibody or nonspecific Rabbit IgG (control) to pulldown endogenous PI3 Kinase p85 ( n = 3 biologically independent samples). (C) Representative immunofluorescence analysis of JUP in control and Optn KD C2C12 cells transfected with Tdtomato-JUP plasmids ( n = 3 biologically independent samples). Scale bars: 5 μm. (D, E) The cytosol and membrane fraction levels of JUP and PI3 Kinase p85 in Optn KD (D) or overexpressing (E) C2C12 cells by immunoblotting analysis ( n = 3 biologically independent samples). The Original blot for this figure can be found in .

Article Snippet: To verify the activation of PI3K-AKT signaling pathway in vivo, the TA muscle of scrambled shRNA or sh Optn was injected with 20 ul the specific PI3K activator 740-YP (30 μM, HY-P0175, MedChem Express, Shianghai, PRC) or DMSO per mouse for 4 weeks.

Techniques: Immunoprecipitation, Incubation, Control, Immunofluorescence, Transfection, Membrane, Western Blot

Journal: PLOS Biology

Article Title: Optineurin binding to the novel interacting partner Junction plakoglobin prevents muscle atrophy in mice

doi: 10.1371/journal.pbio.3003581

Figure Lengend Snippet: Left panel. In the presence of OPTN, it binds to JUP and coordinates the interaction between PI3 Kinase p85 and JUP in normal skeletal muscle, promoting activation of the PI3K-AKT pathway. Right panel. OPTN deficiency decreases the binding between PI3 Kinase p85 and JUP, leading to down-regulation of PI3K-AKT pathway. Consequently, the expression levels of Atrogin-1 and Murf-1 were increased, promoting protein breakdown and muscle atrophy. IGFR, insulin-like growth factor receptor; IR, insulin receptor; IRS, insulin receptor substrate; OPTN, optineurin.

Article Snippet: To verify the activation of PI3K-AKT signaling pathway in vivo, the TA muscle of scrambled shRNA or sh Optn was injected with 20 ul the specific PI3K activator 740-YP (30 μM, HY-P0175, MedChem Express, Shianghai, PRC) or DMSO per mouse for 4 weeks.

Techniques: Activation Assay, Binding Assay, Expressing