Journal: Scientific Reports

Article Title: Hydrogen Sulphide Treatment Increases Insulin Sensitivity and Improves Oxidant Metabolism through the CaMKKbeta-AMPK Pathway in PA-Induced IR C2C12 Cells

doi: 10.1038/s41598-017-13251-0

Figure Lengend Snippet: Information about the antibodies.

Article Snippet: IRS1 (phospho Ser-312) , IRS1 (phospho Ser 312) antibody , GeneTex; GTX24865 , Rabbit polyclonal , 1:1000.

Techniques:

Journal: Scientific Reports

Article Title: Hydrogen Sulphide Treatment Increases Insulin Sensitivity and Improves Oxidant Metabolism through the CaMKKbeta-AMPK Pathway in PA-Induced IR C2C12 Cells

doi: 10.1038/s41598-017-13251-0

Figure Lengend Snippet: Exogenous NaHS promoted glucose uptake and increased the phosphorylation of insulin signalling pathway components in C2C12 cells. After insulin resistance was induced by PA, culture medium was completely removed and replaced with NaHS solution. NaHS solution was freshly prepared and diluted to 50 μM with DMEM. ( A ) Cells were incubated in NaHS solution for 30 minutes at 37 °C, followed by incubation with 10 nM insulin and 150 μg/ml 2-NBDG. NaHS treatment promoted glucose uptake in both control C2C12 cells and the IR cell model. ( B ) Phosphorylation levels of IRS1, PI3K and AKT were evaluated by Western blot. In the PA-treated group, phosphorylation of IRS1, PI3K and AKT were decreased. Administration of NaHS increased the phosphorylation of IRS1, PI3K and AKT. *p < 0.05, **p < 0.01, ***p < 0.001. Data are shown as the mean ± sem.

Article Snippet: IRS1 (phospho Ser-312) , IRS1 (phospho Ser 312) antibody , GeneTex; GTX24865 , Rabbit polyclonal , 1:1000.

Techniques: Phospho-proteomics, Incubation, Control, Western Blot

Journal: Scientific Reports

Article Title: Hydrogen Sulphide Treatment Increases Insulin Sensitivity and Improves Oxidant Metabolism through the CaMKKbeta-AMPK Pathway in PA-Induced IR C2C12 Cells

doi: 10.1038/s41598-017-13251-0

Figure Lengend Snippet: Compound C decreases insulin signalling and reduced NaHS-induced glucose uptake in IR cells. PA-treated C2C12 cells incubated with compound C (20 μM, 30 min) were cultured with DMEM containing 50 μM NaHS for 30 min. ( a ) NaHS-induced phosphorylation of IRS1, PI3K, AKT and AMPK was blocked by compound C. ( b ) NaHS-induced enhancement of glucose uptake was inhibited by compound C (20 μM, 30 min) in IR cell models. *p < 0.05, **p < 0.01, ***p < 0.001. Data are shown as the mean ± sem.

Article Snippet: IRS1 (phospho Ser-312) , IRS1 (phospho Ser 312) antibody , GeneTex; GTX24865 , Rabbit polyclonal , 1:1000.

Techniques: Incubation, Cell Culture, Phospho-proteomics

Journal: Scientific Reports

Article Title: Hydrogen Sulphide Treatment Increases Insulin Sensitivity and Improves Oxidant Metabolism through the CaMKKbeta-AMPK Pathway in PA-Induced IR C2C12 Cells

doi: 10.1038/s41598-017-13251-0

Figure Lengend Snippet: STO-609 inhibition of the effect of NaHS to stimulate the insulin signalling pathway and AMPK to improve glucose uptake in the IR cell model. ( a ) C2C12 myoblasts were treated with the CaMKKβ inhibitor, STO-609 (25 μM, 30 min), and stimulated with or without insulin (10 nM, 10 min). STO-609 blocked the phosphorylation of IRS1, PI3K, AKT and AMPK induced by NaHS. ( b ) Additional increases of the NaHS-induced 2-NBDG uptake was blocked by STO-609 (25 μM, 30 min) in PA-induced IR C2C12 cells. **p < 0.01, ***p < 0.001. Data are shown as the mean ± sem.

Article Snippet: IRS1 (phospho Ser-312) , IRS1 (phospho Ser 312) antibody , GeneTex; GTX24865 , Rabbit polyclonal , 1:1000.

Techniques: Inhibition, Phospho-proteomics

Journal: Virology

Article Title: Molecular pathways for glucose homeostasis, insulin signaling and autophagy in hepatitis C virus induced insulin resistance in a cellular model.

doi: 10.1016/j.virol.2012.07.003

Figure Lengend Snippet: Fig. 6. Expression of autophagic markers. Detection and quantification of autophagic marker proteins in HCV infected cells by Western blot analysis. Equal amounts of total protein (50 mg) in whole cell lysate at each time point was fractionated and analyzed, as described in the Materials and methods section. Fold change with respect to the internal control is indicated beneath each lane. (A) Beclin 1 (62 kda) at two different time points (B) Conjugate of Atg5/Atg12 (53 kda) (using Atg5 antibody) at different time points of post culturing; (C) conjugated complex of Atg5/Atg12 at 48 h recognized individually by Atg5 and Atg12 antibody at two different protein concentrations. Blots of the whole gels are shown in the Supplemental data (Figs. 6D and E).

Article Snippet: Mouse, rabbit or goat antibodies to IRS-1, Atg5, Atg12, GS and actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA; Cell Signaling, Danvers, MA); IRS-1 Ser312 from Abcam (Cambridge, MA); and Beclin1, GSK-3a, GSK-3b, GSK-3aSer21, GSK-3bSer9, IRS-1 Ser302, Ser612, Ser789 and GS Ser641, Atg5, Atg12 and tubulin from cell signaling (Danvers, MA) including conformation specific secondary antibody and anti-mouse HRP for detecting secondary antibody in IP experiments.

Techniques: Expressing, Marker, Infection, Western Blot, Control