egf neutralizing antibody  (Sino Biological)

Journal: PLOS ONE

Article Title: Crystalline silica-exposed human lung epithelial cells presented enhanced anchorage-independent growth with upregulated expression of BRD4 and EZH2 in autocrine and paracrine manners

doi: 10.1371/journal.pone.0285354

Figure Lengend Snippet: (A) ELISA of EGF, TNF-α, IL-6, and TGF-β1 was performed with the culture supernatants of nonadherent NL20, BEAS-2B, and 16HBE 4–6 days after incubation with autocrine CS or BPDE CM compared with the autocrine control CM. ELISA of EGF, TNF-α, and IL-6 was also conducted with the culture supernatants 48 h after exposure to CS in adherent bronchial cell lines. Asterisks indicate a significant difference in concentration (*p < 0.05, **p < 0.01). Relative values are shown as mean ± SD from three independent experiments. (B) Anchorage-independent growth of NL20, BEAS-2B, and 16HBE cells was measured using MTT 5–6 days after the administration of recombinant human EGF, TNF-α, and IL-6. The MTT assay was performed for autocrine CS CM-incubated nonadherent NL20 and BEAS-2B cells four and five days after EGF-neutralizing antibody (NtAb) treatment, respectively, and for autocrine CS CM-incubated nonadherent 16HBE cells, five days after TNF-α NtAb treatment. The MTT assay was also performed for autocrine CS CM-incubated nonadherent bronchial cell lines five days after IL-6 NtAb administration. Asterisks indicate a significant difference in growth (*p < 0.05, **p < 0.01). Relative values are shown as mean ± SD from three independent experiments. (C) Expression of BRD4 and EZH2 in nonadherent 16HBE cells three days after the administration of recombinant human TNF-α determined using western blotting and the expression of BRD4 and EZH2 3–4 days after treatment of nonadherent NL20 and BEAS-2B cells with recombinant human EGF.

Article Snippet: Mouse anti-human EGF neutralizing antibody (NtAb), goat anti-human TNF-α NtAb, and mouse anti-human IL-6 NtAb were obtained from R&D Systems.

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Control, Concentration Assay, Recombinant, MTT Assay, Expressing, Western Blot

Journal: American Journal of Cancer Research

Article Title: Induction of cellular senescence in fibroblasts through β1-integrin activation by tenascin-C-derived peptide and its protumor effect

doi:

Figure Lengend Snippet: TNIIIA2 secreted SASP factors, including HB-EGF, induce the malignant transformation of HaCaT cells. In (A and B), HaCaT cells cocultured with TNIIIA2-induced senescent WI-38 cells were incubated with either an anti-HB-EGF neutralizing antibody or isotype control. After incubation for 72 h, the HaCaT area was measured by rhodanile blue staining. Scale bar, 200 μm. (C) HaCaT cells treated with CM from TNIIIA2-induced senescent WI-38 cells were incubated with either an anti-HB-EGF neutralizing antibody or isotype control and then subjected to a WST-8 (D) or colony formation (E) assay. (F) WI-38 cells were treated with either TNIIIA2 (50 μg/mL) or NaB (4 mM) to induce cellular senescence, as described in the Materials and Methods. HB-EGF and GAPDH mRNA levels were then measured by semi-quantitative PCR. In (G and H), HaCaT cells were treated with recombinant human HB-EGF, and the cells were subjected to a WST-8 assay on day 2 (G) or colony formation assay on day 10 (H). Data are presented as means ± SD, *P < 0.05, **P < 0.01.

Article Snippet: Recombinant human heparin-binding epidermal growth factor-like growth factor (HB-EGF) and an anti-HB-EGF neutralizing antibody (AF-259-NA) were obtained from R&D Systems (Minneapolis, MN).

Techniques: Transformation Assay, Incubation, Control, Staining, Real-time Polymerase Chain Reaction, Recombinant, Colony Assay

Journal: American Journal of Cancer Research

Article Title: Induction of cellular senescence in fibroblasts through β1-integrin activation by tenascin-C-derived peptide and its protumor effect

doi:

Figure Lengend Snippet: A model of malignant transformation of preneoplastic cells via factors secreted from TNIIIA2-induced senescent fibroblasts. Activation of β1-integrin by TNIIIA2 induces fibroblast senescence. Factors secreted by TNIIIA2-induced senescent fibroblasts, including HB-EGF, are involved in the malignant transformation of preneoplastic cells. Inactivation of β1-integrin by peptide FNIII14 inhibits TNIIIA2-induced cellular senescence. Created using BioRender.com.

Article Snippet: Recombinant human heparin-binding epidermal growth factor-like growth factor (HB-EGF) and an anti-HB-EGF neutralizing antibody (AF-259-NA) were obtained from R&D Systems (Minneapolis, MN).

Techniques: Transformation Assay, Activation Assay