Journal: Cancer cell international

Article Title: The association between Salt-inducible kinase 2 (SIK2) and gamma isoform of the regulatory subunit B55 of PP2A (B55gamma) contributes to the survival of glioma cells under glucose depletion through inhibiting the phosphorylation of S6K.

doi: 10.1186/s12935-015-0164-6

Figure Lengend Snippet: Figure 3 Overexpression of B55gamma in Glioma cells inhibits the phosphorylation of S6K. A. Overexpression of B55gamma in U251 decreases phosphorylation of S6K, but phosphorylation of 4EBP1 and mTOR and AKT have no significant change. B. U251 cells were transfected with control vector or B55gamma alone or B55gamma + S6K WT or B55gamma + S6K KA for 48 hrs. Cell viability assays were performed under the treatments of Rapamycin at 15 nM and 50 nM, DMSO treatment was control. C. Transfection of S6K WT and S6K KA into U251V and U251B55gamma cells followed by the measurements of cell viabilities under the glucose withdrawal for 24 hrs. Columns, mean of three independent experiments; bars, SE. **, P < 0.01; ***, P < 0.001.

Article Snippet: Vector containing GFPtagged ORF clone of Homo sapiens protein B55gamma, WT S6K or KA S6K were purchased from Origene.

Techniques: Over Expression, Phospho-proteomics, Transfection, Control, Plasmid Preparation

Journal: Cancer cell international

Article Title: The association between Salt-inducible kinase 2 (SIK2) and gamma isoform of the regulatory subunit B55 of PP2A (B55gamma) contributes to the survival of glioma cells under glucose depletion through inhibiting the phosphorylation of S6K.

doi: 10.1186/s12935-015-0164-6

Figure Lengend Snippet: Figure 5 Knockdown of SIK in B55gamma overexpression cells recovers the phosphorylation of S6K. A. U251V, U251B55gamma, U138V and U138B55gamma cells were transfected with scramble siRNA (Ctr) or SIK2 siRNA. 48 hrs after siRNA transfection, cell lysates were prepared and Western blotting was performed to detect the phosphorylation of S6K statues. The β-actin protein was used as a loading control. B. U251B55gamma (left) and U138B55gamma (right) cells were transfected with scramble siRNA (Ctr) or SIK2 siRNA. 48 hrs after siRNA transfection, cell were plated into 6-well plate for overnight, then the regular medium was replaced by glucose free medium for 36 h-starvation followed by the cell viability assay. Columns, mean of three independent experiments; bars, SE. *, P < 0.05.

Article Snippet: Vector containing GFPtagged ORF clone of Homo sapiens protein B55gamma, WT S6K or KA S6K were purchased from Origene.

Techniques: Knockdown, Over Expression, Phospho-proteomics, Transfection, Western Blot, Control, Viability Assay

Journal: Frontiers in Nutrition

Article Title: Maternal docosahexaenoic acid supplementation during lactation improves exercise performance, enhances intestinal glucose absorption and modulates gut microbiota in weaning offspring mice

doi: 10.3389/fnut.2024.1423576

Figure Lengend Snippet: Effect of maternal DHA supplementation during lactation on the mTOR signaling pathway in weaning mice. (A) Representative Western blots of p-mTOR (S2448), total mTOR, p-p70S6 kinase (T389), total p70S6 kinase, p-S6 ribosomal protein (S240/244) and total S6 ribosomal protein, and (B, C, D) the relative band intensities of phosphorylation levels normalized to their respective total proteins in jejunal tissue of mice from different groups. Data were expressed as the mean ± SEM; n = 4 per group. Significance was set at p < 0.05 ( * p < 0.05, ** p < 0.01). L-DHA: 150 mg/(kg body weight day) DHA; H-DHA: 450 mg/(kg body weight day) DHA. DHA, docosahexaenoic acid; mTOR, mammalian target of rapamycin.

Article Snippet: After transfer, the PVDF membrane was blocked for 1 h in 5% fat-free milk and then was, respectively, incubated overnight at 4°C with dilutions of primary antibodies: GLUT2 (A12307, 1:2000, Abclonal, China), SGLT1 (bs-1128R, 1:2000, Bioss, China), S6 ribosomal protein (abs131865, 1:2000, Absin, China), p-Ser240/244-S6 ribosomal protein (ab215214, 1:2000, Abcam, UK), p70S6 kinase (14485-1-AP, 1,1,000, Proteintech, China), p-Thr389-p70S6 kinase (9,234, 1,1,000, Cell Signaling Technology, United States), mammalian target of rapamycin (mTOR, T55306, 1:2000, Abmart, China), p-S2448-mTOR (5,536, 1,1,000, Cell Signaling Technology), β-actin (T40104, 1:4000, Abmart).

Techniques: Western Blot, Phospho-proteomics

Journal: Neuroscience letters

Article Title: The mTOR cell signaling pathway is crucial to the long-term protective effects of ischemic postconditioning against stroke

doi: 10.1016/j.neulet.2018.03.062

Figure Lengend Snippet: Antibodies, concentrations and manufacturers used

Article Snippet: The S6K1 gene was cloned from S6K1 (pcDNA3) plasmids (S6K1: 26610, Addgene, Cambridge, MA) and inserted into the lentiviral backbone plasmid, pHR’trIPostCMV-IRES-eGFP.

Techniques: