Journal: Journal of Functional Foods

Article Title: Antidiabetic effects and mechanisms of action of γ-conglutin from lupin seeds

doi: 10.1016/j.jff.2021.104786

Figure Lengend Snippet: Fig. 4. γ-conglutin peptides stimulate mTOR downstream pathway in HSMM: Western blot analysis of (A) mammalian target of rapamycin (mTOR) and (B) AKT (protein kinase B) (C) glycogen synthase kinase-3β (GSK3β) (D) 70 KDa ribosomal protein S6 kinase (70S6K) (E) ribosomal protein S6 (S6) and (F) extracellular signal-regulated kinase (ERK1/2) in HSMM cell lysate and ratio of their band intensities (phosphorylated (p)/total (t) or phosphorylated (p)/GAPDH) in presence of vehicle (control referred to as ‘C’), positive control – insulin (100 nM) (referred as ‘I’) or γ-conglutin peptides (200–2 µg/mL). The ratio of band intensities is reported as values normalised with control. Values are mean ± standard deviation of three or more independent experiments; ****p ≤0.0001, ***p ≤0.001, **p ≤0.01 and *p ≤0.05 are significantly different as compared to control. Immunoblots shown are representative of three or more independent experiments.

Article Snippet: Polyclonal goat antirabbit immunoglobulins/HRP from Dako (Glostrup, Denmark) and protease/phosphatase inhibitor cocktail (100X) and primary rabbit mAb (p-mTOR (Ser 2448; #2971S), p-AKT (Ser 473, #9271S), p-S6 (Ser235/ 236, #4858S), p-GSK3B (Ser9; 9336S), p-ERK1/2 (#4370), GAPDH (#D16H11), mTOR (#2972S), AKT (#9272S), S6 (#2217S), GSK3B (#9315S), ERK1/2 (#4695)) were obtained from Cell Signaling Technology (Massachusetts, USA).

Techniques: Western Blot, Control, Positive Control, Standard Deviation

Journal: Materials & Design

Article Title: 3D printed porous microgel for lung cancer cells culture in vitro

doi: 10.1016/j.matdes.2021.110079

Figure Lengend Snippet: Fig. 4. (a, b, c) RNA expression levels of ROCK, Limk, and Cofilin genes in LL2 cells cultured in 2D petri dish and 3D porous microgels, respectively. Results are shown as mean ± S.E.M. of three independent experiments. * p 0.05, *** p 0.001; (d, e) Protein expression levels of ROCK1, G-Actin, and GAPDH in LL2 cells cultured in 2D petri dish and 3D porous microgels, respectively. Results are shown as mean ± S.E.M. of three independent experiments. * p 0.05, ** p 0.01.

Article Snippet: The antibodies included ROCK1 Rabbit mAb, Pan-Actin Rabbit mAb and GAPDH Rabbit mAb obtained from Cell Signalling Technology; and Goat Anti-Rabbit IgG H&L (HRP), which were acquired from Abcam (Shanghai, China).

Techniques: RNA Expression, Cell Culture, Expressing

Journal: Journal of Advanced Research

Article Title: Gut microbial GABAergic signaling improves stress-associated innate immunity to respiratory viral infection

doi: 10.1016/j.jare.2023.06.008

Figure Lengend Snippet: GABA acts through the αKG/Tet2/ PPARγ axis to mediate metabolic-epigenetic regulation of AMs. (A) Schematic diagram of the TCA cycle, with the key enzymes regulated upon stress labeled. Red arrow, down-regulated, and Green arrow, up-regulated in stressed AMs versus non-stressed AMs post IAV infection. (B) Serum levels of αKG and succinate analyzed by GC-MS, with the ratio of αKG to succinate provided. (C-F) Stressed and Non-stressed (NS) mice were infected or uninfected with IAV for 3 days. CD45 + CD11b - CD64 + CD11c + SiglecF + AMs were sorted for subsequent analysis. (C) Immunoblotting of Tet2; (D) Dot-blot analysis of 5hmc or 5mc level. MB, methylene blue staining (as loading control). (E) ChIP test of 5hmc or 5mc enrichment at the pparγ locus; (F) Immunoblotting of PPARγ; (G) Immunoblotting of Tet2 and PPARγ in MH-S cells treated with GABA at the indicated doses; (H) Immunoblotting of Tet2 or PPARγ in GABA-treated macrophages with or without bicuculline (Bic; 40 μM). (I-K) MH-S cells were transfected with Tet2 siRNA (siTet2) or siRNA (siNC), and then treated with GABA (100μM) or 0.01% DMSO prior to IAV infection. (I) PPARγ luciferase activity; (J) Immunoblotting of Tet2 and PPARγ; (K) Heatmapping of AM-associated genes. Shown are representative images. The bar graph is the densitometric quantification of western blot bands. Data from three individual experiments are shown as means ± SD. * P < 0.05, ** P < 0.01, with one-way ANOVA (three or more groups) followed by Bonferroni post hoc test.

Article Snippet: Solution chromatin was immunoprecipitated with anti-5hmc (#39069, 2 μg), anti-5mc (#ab10805, 2 μg), or normal rabbit IgG (CST, #2729, 2 μg), respectively.

Techniques: Labeling, Infection, Gas Chromatography-Mass Spectrometry, Western Blot, Dot Blot, Staining, Transfection, Luciferase, Activity Assay