phospho pi3k p pi3k  (Bioss)

Journal: PLOS One

Article Title: Butyrate ameliorates DSS-induced ulcerative colitis in mice by facilitating autophagy in intestinal epithelial cells and modulating the gut microbiota through blocking the PI3K-AKT-mTOR pathway

doi: 10.1371/journal.pone.0337214

Figure Lengend Snippet: (A) (B) Western blot of autophagy-related proteins. (C) Western blot of PI3K/AKT/mTOR pathway proteins. (D–J) Quantitative analysis of (D) Beclin-1, (E) Atg5, (F) LC3II/I, (G) P62, (H) p-PI3K/PI3K, (I) p-AKT/AKT, and (J) p-mTOR/mTOR protein levels. Data are mean ± SEM; #P < 0.05, ##P < 0.01 vs Control; *P < 0.05, **P < 0.01 vs DSS group.

Article Snippet: PI3K, P62, and Beclin-1 antibodies were obtained from ABclonal (A19742, A19700, and A21191, Wuhan, China), and phospho-PI3K (p-PI3K) from Bioss (bs-5570R, Beijing, China).

Techniques: Western Blot, Control

Journal: Bioactive Materials

Article Title: Pathology-inspired collagen-binding thermosensitive micelle drops enable prolonged and efficient treatment of fungal keratitis

doi: 10.1016/j.bioactmat.2025.04.011

Figure Lengend Snippet: Therapeutic mechanism of PX-TA-AmB in FK via MAPK6/PI3K/AKT pathway modulation. (A) Volcano plot of the differentially expressed genes. (B) KEGG pathway enrichment analysis of the differentially expressed genes. (C) Heatmap of the differentially expressed genes related to the inflammatory response, signaling pathway regulation, and corneal scarring before and after PX-TA-AmB treatment. (D, E) GSEA with the indicated KEGG and Reactome gene sets. (F, G, H) MAPK6, α-SMA, and LOX mRNA expression levels in treated versus control corneal tissues. (I) Representative Western blot images of p-AKT, AKT, p-PI3K, PI3K, MAPK6, Vinculin, and β-actin proteins in different treatment groups. (J) Representative Western blot images of IL-1β and MMP9 in different treatment groups. (K) Representative Western blot images for functional validation of the role of AKT pathway in PX-TA-AmB-mediated downregulation of IL-1β and MMP9 using the AKT activator SC79. (L) Immunofluorescence assessment of α-SMA, MMP9, CD206, CD86, and Ly6G under different treatments. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. Mean ± SD, n = 3; one-way ANOVA.

Article Snippet: The total protein was separated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto a 0.45 μm pore size polyvinylidene fluoride (PVDF) membrane (Millipore, Germany); this membrane was then blocked with 5 % milk at room temperature and incubated with primary antibodies overnight at 4 °C; these antibodies included p-PI3K (Cell Signaling Technology, 4228, 1:1000), PI3K (Cell Signaling Technology, 4257, 1:1000), p-Akt (ABclonal, AP1453, 1:1000), Akt (ABclonal, A18675, 1:2000), MAPK6 (Abcam, ab53277, 1:1000), IL-1β (Abcam, ab9722, 1:1000), MMP9 (Proteintech, 10375-2-AP, 1:1000), Vinculin (Proteintech, 26520-1-AP, 1:10000) and β-Actin (ABclonal, AC026, 1:20000).

Techniques: Expressing, Control, Western Blot, Functional Assay, Biomarker Discovery, Immunofluorescence

Journal: Redox Biology

Article Title: Dysregulation of sphingolipid and cholesterol homeostasis imposes oxidative stress in human spermatozoa

doi: 10.1016/j.redox.2025.103669

Figure Lengend Snippet: MβCD increases PI3K phosphorylation, while inhibition of S1PR or NOS impairs MβCD-induced tyrosine phosphorylation. Spermatozoa were incubated for 3.5 h at 37 °C under various conditions: untreated (−), treated with 10 % v/v fetal cord serum ultrafiltrate (FCSu), treated with 0.5 mM methyl-β-cyclodextrin (MβCD), and treated with MβCD in combination with ( a ) 40 μM VPC 23019 (S1PR1/3 inhibitor) or ( b ) l -NAME (Nitric Oxide Synthase inhibitor). ( a ) Immunoblotting analysis revealed that VPC 23019 at 40 μM decreased PI3K phosphorylation (P-PI3K) in spermatozoa incubated with 0.5 mM of MβCD. ( b ) Immunoblotting showed that l -NAME treatment decreased tyrosine phosphorylation (P-Tyr) in sperm samples treated with FCSu and MβCD. The signal intensity for each lane was normalized to the silver-stain optical density value for accurate quantification. α-Tubulin loading controls were performed for direct comparison with the silver stain loading controls. The data represent sperm samples from four different healthy donors (n = 4). Statistical analysis was performed using ANOVA with Tukey's test: ∗p ≤ 0.05, ∗∗p ≤ 0.01, and ∗∗∗p ≤ 0.001.

Article Snippet: Rabbit polyclonal anti-phospho-PI3K (P-PI3K) antibodies were purchased from Cell Signaling ( # 4228) (Beverly, MA, USA).

Techniques: Phospho-proteomics, Inhibition, Incubation, Western Blot, Silver Staining, Comparison