Journal: Molecules

Article Title: Revealing Novel Source of Breast Cancer Inhibitors from Seagrass Enhalus acoroides : In Silico and In Vitro Studies

doi: 10.3390/molecules29051082

Figure Lengend Snippet: ΔG of Molecular docking parameter of identified compounds of Seagrass E. acoroides .

Article Snippet: In vitro analysis of HIF-1α, EGFR tyrosine kinase, and HER2 Expressions was carried out in accordance with the manufacturer’s protocol (HIF-1 alpha Monoclonal Antibody (ESEE122), eBioscienceTM; Human EGFR (Epidermal Growth Factor Receptor) ELISA Kit; Elabscience ® for HER2) and established research experimental guidelines [ ].

Techniques: Control

Journal: Molecules

Article Title: Revealing Novel Source of Breast Cancer Inhibitors from Seagrass Enhalus acoroides : In Silico and In Vitro Studies

doi: 10.3390/molecules29051082

Figure Lengend Snippet: Downregulation of HIF-1A, EGFR tyrosine kinase, and HER2 by Seagrass EAE.

Article Snippet: In vitro analysis of HIF-1α, EGFR tyrosine kinase, and HER2 Expressions was carried out in accordance with the manufacturer’s protocol (HIF-1 alpha Monoclonal Antibody (ESEE122), eBioscienceTM; Human EGFR (Epidermal Growth Factor Receptor) ELISA Kit; Elabscience ® for HER2) and established research experimental guidelines [ ].

Techniques:

Journal: Oncology Letters

Article Title: Effects of Yangzheng Sanjie Decoction-containing serum mediated by microRNA-7 on cell proliferation and apoptosis in gastric cancer

doi: 10.3892/ol.2018.7757

Figure Lengend Snippet: Expression of miR-7 and EGFR. (A) Hematoxylin staining indicating the expression of miR-7 by in situ hybridization (magnification, ×400). (B) Expression of miR-7 for reverse transcription-quantitative polymerase chain reaction. U6 was used as the internal control. (C) Western blot analysis of EGFR. (D) Representative image of EGFR for western blot analysis. β-actin was used as the internal control. *P<0.05. miR-7, microRNA-7; EGFR, epidermal growth factor receptor; CM, Chinese medicine; Con, control; GC, gastric cancer.

Article Snippet: The membranes were blocked with 5% milk in Tris-buffered saline and 0.2% Tween at room temperature for 1 h, prior to being incubated overnight at 4°C with specific primary antibodies against the following: β-actin (dilution, 1:250; cat. no. BM0627; Boster Biological Technology, Wuhan, China) and EGFR (dilution, 1:1,000; cat. no. 4267S; CST Biological Reagents Co., Ltd., Shanghai, China).

Techniques: Expressing, Staining, In Situ Hybridization, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Western Blot

Journal: Drug delivery

Article Title: In vivo antitumor activity of PHT-427 inhibitor-loaded polymeric nanoparticles in head and neck squamous cell carcinoma.

doi: 10.1080/10717544.2024.2449376

Figure Lengend Snippet: Figure 3. NP-427 downregulates EGFR expression in the HNSCC model. (A) EGFR protein expression was determined by ELISA. (B) Fluorescence micro graphs of representative immunostaining of EGFR-Alexa Fluor546 conjugate in tumor tissue (red) and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) (blue) (x40), Scale bar 100 µm. (C) EGFR immunofluorescence intensity was ana lyzed by ImageJ software. Experimental groups: control group (CTR) treated with 0.9% saline solution; unloaded nanoparticles group (NP-Ø) with a concen tration of 1 mg/mL; PHT-427 inhibitor-free at concentrations of 0.5 and 1 mg/ mL group (PHT-0.5 and PHT-1, respectively) and loaded nanoparticles with the PHT-427 inhibitor at the concentrations of 0.5 and 1 mg/mL groups (NP-0.5 and NP-1, respectively) for 21 days. The diagrams for EGFR represent the mean ± SD of ng/mL for a and arbitrary units (AU) for B. p < .05 versus CTR and NP-Ø groups. (n = 32).

Article Snippet: Protein extracts were assayed by a different human enzyme-linked immunosorbent assay (elisa) kits by the following proteins: eGFR was quantified using an elisa kit (catalog number: csB-e12124h; cusabio, houston, Usa); akt(ps473) and total akt levels were measured using abcam elisa kit (catalog number: ab126433; abcam, cambridge, UK); Phospho-PDK1 (s241) and total PDK1 by the elisa Kit (catalog number: ab279889) of abcam (cambridge, UK), and human Pi3K by the elisa Kit (catalog number: csB-e08417h, cusabio, houston, Usa) according to the protocol’s instructions. the plates were assessed on the FlUOstar Omega (BMG labtech, Ortenberg, Germany) to 450 nm. these experiments were performed in triplicate for each samples (N = 31) (ctR group (n = 4); NP-Ø group (n = 5); Pht-0.5 group (n = 5); Pht-1 (n = 5); NP-0.5 (n = 6); and NP-1 group (n = 6)).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Fluorescence, Immunostaining, Immunofluorescence, Software, Control, Saline

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Naringenin suppresses epithelial ovarian cancer by inhibiting proliferation and modulating gut microbiota.

doi: 10.1016/j.phymed.2022.154401

Figure Lengend Snippet: Fig. 4. Naringenin suppressed EOC through the PI3K signaling pathway in vitro. Western blotting analysis of lysates (40 μg) of EOC cells treated with naringenin at the indicated concentrations for 24 h (a, b, i, j). Membranes were incubated with antibodies against EGFR (c, f), PI3K (d, g), CCND1 (e, h) and p-PI3K (k, l). All data are expressed as the mean ± SD of values from experiments performed in triplicate. * p < 0.05; ** p < 0.01; *** p < 0.001 compared to ctrl groups.

Article Snippet: Tissues were stained with primary antibodies against EGFR (1:50, A00023-2, Boster, Wuhan, China), PI3K (1:50, bs-4160R, Bioss, Beijing, China) and CCND1 (1:50, BM4272, Boster, Wuhan, China) for immunohistochemical analysis.

Techniques: In Vitro, Western Blot, Incubation

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Naringenin suppresses epithelial ovarian cancer by inhibiting proliferation and modulating gut microbiota.

doi: 10.1016/j.phymed.2022.154401

Figure Lengend Snippet: Fig. 6. Naringenin suppressed EOC through the PI3K signaling pathway in vivo. HE staining (a) and IHC staining for EGFR (b, f), PI3K (c, g) and CCND1 (d, h) in tumors and quantification (n=5) (Nar p.o. means Nar p.o., per os; ns means not statistically significant, * p < 0.05; ** p < 0.01; *** p < 0.001 compared to untreated; # p < 0.05 Nar p.o., per os compared to Nar i.p.).

Article Snippet: Tissues were stained with primary antibodies against EGFR (1:50, A00023-2, Boster, Wuhan, China), PI3K (1:50, bs-4160R, Bioss, Beijing, China) and CCND1 (1:50, BM4272, Boster, Wuhan, China) for immunohistochemical analysis.

Techniques: In Vivo, Staining, Immunohistochemistry

Journal: Scientific Reports

Article Title: FimH-based display of functional eukaryotic proteins on bacteria surfaces

doi: 10.1038/s41598-019-44883-z

Figure Lengend Snippet: Design and expression of POI-FimH fusion proteins. ( A ) The Gaussia Luciferase (GLuc) is N-linked to the FimH protein and thereby becomes part of the fimbriae on the bacterial surface. By use of the bacterial pet11d vector, the expression of the GLuc-FimH fusion protein is induced by incubation with IPTG. ( B ) Schematic representation of the expression cassettes for the FimH fusion proteins with human EGF, human TGF-α and human epiregulin EPRG, respectively. Gly, Glycine-(Gly-Gly-Gly-Gly-Ser) 3 -linker; cMyc, c-Myc tag; CS, HRV3C protease cleavage site. ( C ) The expression of the POI-FimH protein on the surface of E. coli was induced by incubation with IPTG for 2 hrs and recorded by flow cytometry. Bacteria containing the GLuc-FimH expression vector were stained with a Gaussia luciferase-specific rabbit antibody and detected with a PE-coupled anti-rabbit antibody (bold histogram). An isotype-matched PE-conjugated antibody (light histogram) and non-modified E. coli (wt) served as controls. The expression of EGF-FimH, TGFα-FimH and Epiregulin-FimH on the surface of engineered E. coli bacteria was induced by IPTG. The bacteria were stained with a PE labeled c-Myc-specific mouse antibody and the proteins were recorded by flow cytometry (bold histograms). An isotype-matched PE-conjugated antibody (light histograms) served as control. All experiments were performed in triplicates and a representative histogram is shown.

Article Snippet: The detection of the proteins of interest was performed by using ELISA kits specific for human EGF, TGF-α and EPRG, respectively (CUSABIO BIOTECH, Houston, TX, USA).

Techniques: Expressing, Luciferase, Plasmid Preparation, Incubation, Ubiquitin Proteomics, Flow Cytometry, Bacteria, Staining, Modification, Labeling, Control

Journal: Scientific Reports

Article Title: FimH-based display of functional eukaryotic proteins on bacteria surfaces

doi: 10.1038/s41598-019-44883-z

Figure Lengend Snippet: Recombinant EGFR ligands produced by transformed E. coli induce EGFR receptor phosphorylation. ( A ) E. coli bacteria were engineered with FimH fusion proteins linked with the EGFR ligand epiregulin, EGF or TGF-α, respectively. Displayed EGFR ligands were proteolytically released from the bacteria surface and detected by ELISA. The analysis was performed in triplicates for each condition and the entire experiment repeated at least twice; representative data are shown. ( B ) Proteolytically released proteins of interest, i.e., EGF, TGF-α and epiregulin were tested for binding to the EGF receptor (EGFR). Therefore, microtiter wells were coated with the bacteria produced EGF, TGF-α and epiregulin, respectively, and incubated with the human EGFR-hIgG (filled symbols) and mouse EGFR-IgG (open symbols) fusion protein to test for specific binding. A human IgG fusion protein of irrelevant specificity (bold line) served as control. Bound EGFR-IgG fusion protein was detected by a biotin-labeled human IgG-specific antibody. ( C ) EGFR + 293T cells were incubated in the presence of proteolytically released EGF, TGF-α or EPRG, respectively, and tested for EGFR (Tyr1173) phosphorylation by flow cytometry. The mean fluorescence intensity (MFI) after incubation with the EGFR ligands is shown. All experiments were performed in triplicates for each condition and repeated at least twice; a representative experiment is shown.

Article Snippet: The detection of the proteins of interest was performed by using ELISA kits specific for human EGF, TGF-α and EPRG, respectively (CUSABIO BIOTECH, Houston, TX, USA).

Techniques: Recombinant, Produced, Transformation Assay, Phospho-proteomics, Bacteria, Enzyme-linked Immunosorbent Assay, Binding Assay, Incubation, Control, Labeling, Flow Cytometry, Fluorescence