Journal: Molecular Vision

Article Title: Glucosamine inhibits epidermal growth factor-induced proliferation and cell-cycle progression in retinal pigment epithelial cells

doi:

Figure Lengend Snippet: Effects of glucosamine (GlcN) on transactivation of epidermal growth factor receptor (EGFR). Human retinal pigment epithelial cell line (ARPE-19) cells were treated with 2.5 mM or 5.0 mM GlcN, or 30 mM glucose for 24 h, stimulated with 10 ng/ml EGF, and stained with PE-conjugated anti-total-EGFR antibody and Alexa-Fluor-647-conjugated anti-p-EGFR antibody (Y845). The levels of phosphorylated ( A ) and total EGFR ( B ) were analyzed by flow cytometry. The data are representative of at least three independent experiments. Cells cultured under serum-free conditions were used as the control. C : western blot analysis of ARPE-19 cells cultured to 80% confluence; treated with 2 μg/ml tunicamycin, 30 mM glucose, or 5 mM GlcN in serum-free medium for a further 24 h; then stimulated with EGF for 5 min.

Article Snippet: The following primary antibodies were used: phycoerythrin (PE)-conjugated anti-total-EGFR (cells were not permeabilized for total EGFR detection) and Alexa-Fluor-647-conjugated anti-p-EGFR (Y845; BD PharMingen).

Techniques: Staining, Flow Cytometry, Cell Culture, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Oxyresveratrol as a novel ferroptosis inducer exhibits anticancer activity against breast cancer via the EGFR/PI3K/AKT/GPX4 signalling axis

doi: 10.3389/fphar.2024.1527286

Figure Lengend Snippet: ORes induces ferroptosis in breast cancer cells via the EGFR/PI3K/AKT/GPX4 signalling axis. (A) The table indicates the top 10 connectivity scores between the input gene signature and the gene signatures of compounds in the LINCS L1000 touchstone dataset. (B) The table displays the top 10 scores between the input gene signature and the gene signatures of compounds in the DGDB dataset. (C) GSEA of HTS 2 results of Osimertinib dimesylate–treated MDA-MB-231 cells. (D) GSEA of HTS 2 results of AZ-5104–treated MDA-MB-231 cells. (E) Volcano plot of the DEGs in EGFR knockdown MDA-MB-231 cells, with red and blue dots indicating upregulated and downregulated genes, respectively. Differential gene screening criteria: |FoldChange|>1.3 and p- value < 0.05. (F) Representative Western blot results. (G) Quantification of western blots. Experiments were performed in triplicate, and data are presented as mean ± SD. *** p < 0.001; ns, no significance.

Article Snippet: The membranes were incubated overnight at 4°C with primary antibodies against GPX4 (Cell Signaling Technology, 59735, 1:1,000), phosphorylated EGFR (ZenBio, R26283, 1:1,000), EGFR (Selleck, A5858, 1:1,000), phosphorylated PI3K (ZenBio, 310164, 1:1,000), PI3K (ZenBio, 200900, 1:1,000), phosphorylated AKT (Cell Signaling Technology, 13038, 1:1,000), and AKT (Cell Signaling Technology, 4691, 1:1,000).

Techniques: Knockdown, Western Blot