Journal: Ecotoxicology and environmental safety

Article Title: Azadirachtin exposure inhibit ovary development of Spodoptera litura (Lepidoptera: Noctuidae) by altering lipids metabolism event and inhibiting insulin signaling pathways.

doi: 10.1016/j.ecoenv.2023.115151

Figure Lengend Snippet: Fig. 6. AZA exposure led to insulin resistance- like phenotypes in ovary of S.litura. (A) INS level in ovary. (B) Glu level in ovary. (C) Gly level in ovary. (D) FFA level in ovary. (E) Western blot for insulin signal-related proteins (PI3K, IRS1, P-IRS1 (ser307), IRS2, P-IRS2 (ser731), AKT and P-AKT). (F) The bar graph showed the relative protein abundances fol lowed western blot by normalizing the signals to β-Actin via Image J software. Data was rep resented as mean ± SEM (n = 3), significance was determined by Student’s unpaired t tests. Values significantly differ at * ** p-value < 0.001.

Article Snippet: The membranes were blocked with 5 % skim milk in TBST buffer (Tris-HCl, pH 7.5, 150 mM NaCl, and 0.1 % Tween 20) for 2 h. After washing three times with PBST, the membranes were incubated with primary antibodies against the following proteins, IRS1 (PB0221, Boster, Wuhan, China), P-IRS1 (ser307) (abs130899, Absin, Shanghai, China), IRS2 (PB0222, Boster, Wuhan, China), P-IRS2 (ser731) (bs-10998R, Bioss, Beijing, China), AKT (bs-6951R, Bioss, Beijing, China), P-AKT (ser505) (#4054, Cell Signaling Technology, USA), PI3K p85 (#4257, Cell Signaling Technology, USA) and the reference proteins (β-Actin) (AF003, Beyotime, Shanghai, China), respectively.

Techniques: Western Blot, Software

Journal: Molecular Medicine

Article Title: Induction of Steatohepatitis (NASH) with Insulin Resistance in Wild-type B6 Mice by a Western-type Diet Containing Soybean Oil and Cholesterol

doi: 10.2119/molmed.2016.00203

Figure Lengend Snippet: Dietary cholesterol in soybean oil diet impaired hepatic insulin signaling. Mice received diets as described in Figure 1. Protein levels of (A) insulin receptor substrate 1 (IRS-1) and (B) IRS-2 were determined in liver lysates by Western blot analysis with specific antibodies. Density units were normalized to Fast Green staining, which was verified on the same Western blot membrane as a loading control. (C) Hepatic glycogen content was enzymatically quantified in homogenates (see Materials and Methods section). Values represent means ± standard error of the mean of 5 mice per group. Statistics: One-way analysis of variance with Dunnett’s post hoc test or, where appropriate, Tukey’s post hoc test for multiple comparisons, *p <0.05.

Article Snippet: Western Blot Analysis and Oxyblot Western blot was performed as previously described ( 20 ) with anti-Prx-SO 3 (Abcam, Cambridge, UK), anti-IRS-1 and anti-IRS-2 antibodies (NEB, Frankfurt am Main, Germany) as well as FastGreen staining (Sigma-Aldrich) as a loading control.

Techniques: Western Blot, Staining