intracellular phosphorylated s6 ribosomal protein ps6 levels  (Cell Signaling Technology Inc)

Journal: Cell proliferation

Article Title: The apoptotic and anti-proliferative effects of Neosetophomone B in T-cell acute lymphoblastic leukaemia via PI3K/AKT/mTOR pathway inhibition.

doi: 10.1111/cpr.13773

Figure Lengend Snippet: FIGURE 5 NSP-B's effect on Protein Kinase B/mammalian target of rapamycin (AKT/MTOR) signalling and apoptotic inhibitors in T cells. (A, B, E) Modulation of AKT/MTOR Pathway by NSP-B: Here, the impact of NSP-B on AKT (Ser473), MTOR (S2448), 4EBP1 and GSK3α/β phosphorylation and the expression of apoptotic inhibitors (XIAP, c-IAP1 and c-IAP2) is shown. Proteins from treated cells were resolved by SDS–PAGE and analysed by immunoblotting using specific antibodies. GAPDH and HSP60 were used as loading controls. (C) mTOR Activity Assessment: This panel depicts the intracellular levels of phosphorylated S6 ribosomal protein (pS6), a downstream target of mTOR, in cells treated with or without 2 μM NSB-P. Anti-pS6 monoclonal antibody was used for intracellular staining, with FACS histograms showing the mean fluorescence intensity (MFI). (D) Statistical analysis of pS6 MFI: A representative plot illustrates the pS6 MFI (mean ± SD) from two independent experiments, each conducted in triplicates. Statistical significance was calculated using an unpaired t-test (****p < 0.0001). NSP-B, Neosetophomone B.

Article Snippet: To assess mTOR activity, intracellular phosphorylated S6 ribosomal protein (pS6) levels were detected as described previously.28 Jurkat cells treated with NSP-B were fixed, permeabilized and stained with anti-pS6RP-Alexa Fluor 647 antibody (Ser235/236, clone 2F9, Cell Signaling Technologies).

Techniques: Phospho-proteomics, Expressing, SDS Page, Western Blot, Activity Assay, Staining, Fluorescence

Journal: Nature Communications

Article Title: X chromosome dosage drives statin-induced dysglycemia and mitochondrial dysfunction

doi: 10.1038/s41467-024-49764-2

Figure Lengend Snippet: Mice were fed chow or statin-containing diet with daily administration of either coconut oil vehicle or fish oil (as a source of ω-3 fatty acids) dissolved in coconut oil vehicle. After 5 weeks, a glucose tolerance and b grip strength were measured in female and male mice ( N = 5 mice/sex on each treatment). AUC, area under the curve for glucose tolerance test. Tissues from mice in a , b (5 mice/sex on each treatment) were used to determine c liver and muscle glycogen levels by biochemical assay, and d levels of total and phosphorylated glycogen synthase kinase−3β (GSK−3β) by immunoblot. The same blot was used sequentially for detection of phosphorylated and total GSK−3β and GAPDH. Values represent mean ± SEM. Data analyses were as in Fig. with p values representing pair-wise two-sided t-tests following significant 2-way ANOVA. Source data are provided as a Source Data file.

Article Snippet: Total and phosphorylated glycogen synthase kinase (GSK) 3β protein levels were determined by immunoblot using antibodies (1:1000 dilution) that recognize phospho-GSK-3β Ser9 (Cell Signaling Technology rabbit polyclonal antibody #9336, lot 14) or total GSK-3β protein (Cell Signaling Technology monoclonal antibody #9315, clone 27C10).

Techniques: Western Blot

Journal: Nature Communications

Article Title: Mechanistic characterization of a Drosophila model of paraneoplastic nephrotic syndrome

doi: 10.1038/s41467-024-45493-8

Figure Lengend Snippet: A – D UMAP showing the gene expression changes of JNK/Jra pathway genes in MT of EGT > w1118 and EGT > yki act flies. E , F qPCR of Jra , kay and puc genes in MT of EGT > yki act and PC ts > Pvr act flies. Transgenes were induced for 8 days. Data are presented as means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 with student t-test. G – J Western blots showing the protein levels of JNK and phosphorylated JNK in MT of EGT > w1118 and EGT > yki act G and PC ts > w1118 and PC ts > Pvr act I flies. H shows the signal in the stem cell zone (SCZ) and principal cells plus stellate cells (PC + SC). J shows the rescue of p-JNK upon knockdown of Pvr in principal cells in yki act flies. Transgenes were induced for 5 and 8 days in G , I ; 8 days in H ; 4 and 7 days in J . n = 3 biologically independent experiments with 2 or 3 technical replicates in E , F .

Article Snippet: G – J Western blots showing the protein levels of JNK and phosphorylated JNK in MT of EGT > w1118 and EGT > yki act G and PC ts > w1118 and PC ts > Pvr act I flies.

Techniques: Gene Expression, Western Blot, Knockdown

Journal: Cancer Research Communications

Article Title: HER2-selective and reversible tyrosine kinase inhibitor tucatinib potentiates the activity of T-DM1 in preclinical models of HER2-positive breast cancer

doi: 10.1158/2767-9764.crc-23-0302

Figure Lengend Snippet: FIGURE 6 The combined suppression of HER2 signaling by tucatinib and T-DM1 is associated with reduced tumor growth. A, ELISAs quantifying phosphorylation of signaling components downstream of HER2 in BT-474 cells (as percentage phosphorylated protein vs. untreated cells with SEMs). B, IHC images of pHER2, pHER3, pAKT, and pMEK staining of the BT-474 xenograft tumor model. Inlay images represent HER2-stained tumor

Article Snippet: Phosphorylated protein levels were normalized to total protein levels (ErbB2, #7310;HER3, #7888C; AKT, #7170C; MEK, #7165C all from CST) and quantified relative to untreated cells.

Techniques: Phospho-proteomics, Staining