Journal: PeerJ

Article Title: Dihydrotanshinone I inhibits hepatocellular carcinoma cells proliferation through DNA damage and EGFR pathway

doi: 10.7717/peerj.15022

Figure Lengend Snippet: (A) Calculated binding mode of dihydrotanshinone I with EGFR. Three-dimensional (3D) presentation of the binding mode. (B) Two-dimensional (2D) presentation of hydrophobic interactions between amino acid residues and dihydrotanshinone I. (C) RMSD of the EGFR- dihydrotanshinone I and the EGFR-EAI045 complex. (D) RMSF of the EGFR- dihydrotanshinone I complex. (E, F) Western blot analysis of the inhibitory effects of dihydrotanshinone I on EGFR and its downstream proteins (AKT and STAT3). All experiments were repeated three times independently.

Article Snippet: Samples were followed incubated with anti-p-EGFR (CST, D38B1, 1:1000), anti-EGFR (CST, D7A5, 1:1000), anti-p-STAT3 (CST, D3A7, 1:1000), anti-STAT3 (CST, 124H6, 1:1000), anti-p-AKT (CST, 11E7, 1:1000), anti-BAX (CST, 41162S, 1:1000), anti-Bcl2 (CST, 15071S, 1:1000) and anti-AKT (Proteintech, 66444, 1:1000) primary antibodies at 4 °C overnight.

Techniques: Binding Assay, Western Blot

Journal: BioMed Research International

Article Title: Integrating Network Pharmacology and Experimental Validation to Elucidate the Mechanism of Yiqi Yangyin Decoction in Suppressing Non-Small-Cell Lung Cancer

doi: 10.1155/2023/4967544

Figure Lengend Snippet: Molecular docking. (a–c) Molecular docking of luteolin and AKT1, EGFR, and TP53. (d–f) Molecular docking of quercetin and AKT1, EGFR, and TP53.

Article Snippet: Primary antibodies against p53, p21, cyclin D1, Bax, Bcl-2, cleaved caspase-3, EGFR, p-EGFR, PI3K, p-PI3K, AKT, p-AKT, and GAPDH were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques:

Journal: BioMed Research International

Article Title: Integrating Network Pharmacology and Experimental Validation to Elucidate the Mechanism of Yiqi Yangyin Decoction in Suppressing Non-Small-Cell Lung Cancer

doi: 10.1155/2023/4967544

Figure Lengend Snippet: YYD inactivates EGFR-PI3K-AKT signaling in NSCLC cells. (a, b) Effects of YYD (0, 125, and 250 μ g/ml) on the expression levels of p-EGFR, EGFR, p-PI3K, PI3K, p-AKT, and AKT were determined with western blot assays. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus the nontreated group.

Article Snippet: Primary antibodies against p53, p21, cyclin D1, Bax, Bcl-2, cleaved caspase-3, EGFR, p-EGFR, PI3K, p-PI3K, AKT, p-AKT, and GAPDH were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Western Blot

Journal: BioMed Research International

Article Title: Integrating Network Pharmacology and Experimental Validation to Elucidate the Mechanism of Yiqi Yangyin Decoction in Suppressing Non-Small-Cell Lung Cancer

doi: 10.1155/2023/4967544

Figure Lengend Snippet: EGFR-PI3K-AKT signaling is responsible for the anticancer activity of YYD in NSCLC. Cells were treated with 250 μ g/ml YYD, 100 μ M NSC228155, or simultaneously treated with 250 μ g/ml YYD and 100 μ M NSC228155. (a) Western blot assays were performed to examine the related protein expression. (b–d) Cell proliferation capability was determined. (e, f) Cell cycle distribution and apoptosis were examined. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus the control or YYD group.

Article Snippet: Primary antibodies against p53, p21, cyclin D1, Bax, Bcl-2, cleaved caspase-3, EGFR, p-EGFR, PI3K, p-PI3K, AKT, p-AKT, and GAPDH were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Activity Assay, Western Blot, Expressing

Journal: BioMed Research International

Article Title: Integrating Network Pharmacology and Experimental Validation to Elucidate the Mechanism of Yiqi Yangyin Decoction in Suppressing Non-Small-Cell Lung Cancer

doi: 10.1155/2023/4967544

Figure Lengend Snippet: YYD inhibits NSCLC tumor growth. A549 cells were subcutaneously implanted into nude mice to establish the transplantation tumor models of NSCLC. After 6 days, mice were intragastrically administered with YYD daily. (a) Tumor growth was examined every 3 days. (b) Tumor weight. (c) The expression of proteins associated with EGFR-PI3K-AKT signaling. (d) IHC staining for p-EGFR, Ki-67, cyclin D1, and Bax in xenograft tumors. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus the control group.

Article Snippet: Primary antibodies against p53, p21, cyclin D1, Bax, Bcl-2, cleaved caspase-3, EGFR, p-EGFR, PI3K, p-PI3K, AKT, p-AKT, and GAPDH were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Transplantation Assay, Expressing, Immunohistochemistry

Journal: Biomolecules & Therapeutics

Article Title: Picropodophyllotoxin Inhibits Cell Growth and Induces Apoptosis in Gefitinib-Resistant Non-Small Lung Cancer Cells by Dual-Targeting EGFR and MET

doi: 10.4062/biomolther.2022.113

Figure Lengend Snippet: Effects of PPT on EGFR or MET mediated signaling pathways. Cells were treated with PPT at the indicated concentration for 48 h, then protein expression levels were determined. HCC827GR cell lysates were subjected to western blotting to detect of p-EGFR (Tyr1068), EGFR, p-MET (Tyr1234/1235), MET, p-AKT (Ser473), AKT, p-ERK (Thr202/Tyr204) and ERK antibodies.

Article Snippet: Antibodies against p-EGFR (Tyr1068), EGFR, p-MET (Tyr1234/1235), MET, p-AKT (Ser473), AKT, p-ERK (Thr202/Tyr204), and ERK were purchased from Cell Signaling Biotechnology (Beverly, MA, USA).

Techniques: Concentration Assay, Expressing, Western Blot

Journal: Clinical Epigenetics

Article Title: Phospholipase C delta 1 inhibits WNT/β‐catenin and EGFR-FAK-ERK signaling and is disrupted by promoter CpG methylation in renal cell carcinoma

doi: 10.1186/s13148-023-01448-2

Figure Lengend Snippet: PLCD1 ectopic expression inhibited oncogenic signaling. a Experimentally expressed PLCD1 in 293 T cells was confirmed by Western blot. b Activity of several oncogenic signaling reporters were evaluated using dual-luciferase reporters assay in 293 T cells. b TCF activity and CCND1 , c-Myc and MMP7 promoter reporter activities in PLCD1 -overexpressing A498 (upper) and HH244 (bottom) cells. d Western blot analyses of total β-catenin, p-β-catenin (Ser552), c-Myc, active β-catenin, cyclinD1, MMP7, PLCD1 and GAPDH. e Western blot analyses of EGFR, ERK1/2, Src, FAK and PLCD1 , with GAPDH as internal control. Histograms indicated quantification of the phosphorylated protein normalized to corresponding total protein level. f Schematic diagram of roles and mechanisms of PLCD1 in RCC tumorigenesis

Article Snippet: Antibodies used are: anti-mouse IgG-HRP (DAKO, P0161), anti-rabbit IgG-HRP (DAKO, P0448), GAPDH (Millipore, MAB374), Anti-V5-Tag (Invitrogen, R96025), E-cadherin (CST, 4065), vimentin (Sigma: V6630), Twist (Santa Cruz, sc-15393), MMP7 (Thermo Fisher, MS-813-P0), Total EGFR (CST: 54,359), p-EGFR (CST: 3777), Total FAK (CST: 71,433), p-FAK (Tyr397) (CST: 8556), Total SRC (CST: 2191), p-SRC (CST: 59,548), Total β-catenin (CST: 59,548), active β-catenin (Millipore, 05,665), p-β-catenin (Ser552) (CST: 5651), c-Myc (CST: 18,583), cyclinD1 (CST: 55,506), p-ERK1/2 (CST: 9101), Total ERK1/2 (CST: 4695), Cleaved-Caspase3 (CST: 9661), Cleaved-PARP (CST: 9541), Caspase 3 (CST:9504), PARP (CST: 9532), Bax (CST:2772), Bcl-2 (CST: 2872).

Techniques: Expressing, Western Blot, Activity Assay, Luciferase