Journal: Oncotarget

Article Title: The HGF inhibitory peptide HGP-1 displays promising in vitro and in vivo efficacy for targeted cancer therapy

doi:

Figure Lengend Snippet: a. K D of HGP-1 binding to HGF was determined by SPR technique. b. The assessment of binding competition between various proteins and HGF by fluorescence-based ELISA assay post 1.5-hour incubation. Proteins at the concentrations of 0.05 nM, 0.5 nM, 5 nM and 50 nM mixed with 10 μM FITC-labeled HGP-1 were the liquid phase ( n = 5). c. The binding activity between HGP-1 to HGF and EGF were measured by fluorescence-based direct ELISA assay post 1.5-hour incubation. HGP-1 at the concentrations of 0.1 μM, 1 μM, 10 μM, 100 μM were used ( n = 3). Values were mean ± SEM.

Article Snippet: Recombinant human full length HGF (10463- NHAC-A), VEGF-165 (11066-HNAB), Protein G (13103-PNAE), Biotinylated recombinant human full length HGF (10463-HNAC-B), HGF antibody (10463-RP01-B), MET antibody (10692-MM02), EGF antibody (0605-R008) were purchased from Sino Biological Inc. Recombinant human EGF (H6000–10), bFGF (H3000–10) were obtained from Beijing Wishbiotechnology Co., Ltd. Dynabeads MyOne streptavidin C1 magnetic beads (#65002) and Streptavidin Phycoerythrin Conjugates (S-866) were obtained from Invitrogen.

Techniques: Binding Assay, Fluorescence, Enzyme-linked Immunosorbent Assay, Incubation, Labeling, Activity Assay, Direct ELISA

Journal: Oncotarget

Article Title: The HGF inhibitory peptide HGP-1 displays promising in vitro and in vivo efficacy for targeted cancer therapy

doi:

Figure Lengend Snippet: The cells were treated with HGP-1 (5 μM and 50 μM) and SP-H1 (50 μM) for 15 minutes respectively. a. Representatives Western Blot of phospho-MET level in A549 cells after treated by HGF alone or combining with HGP-1. b. Quantitation of HGP-1 inhbitory effect on phospho-MET level in A549 cells. c. Representatives Western blot of phospho-ERK1/2 level in A549 cells after HGP-1 treatment. d. Quantitaion of HGP-1 inhibitory effect on phospho-ERK1/2 level in A549 cells. * P < 0.05, **0.01 < P < 0.05, represented the significance between HGF and HGP-1 or SP-H1 group. Values were mean ± SEM ( n = 5).

Article Snippet: Recombinant human full length HGF (10463- NHAC-A), VEGF-165 (11066-HNAB), Protein G (13103-PNAE), Biotinylated recombinant human full length HGF (10463-HNAC-B), HGF antibody (10463-RP01-B), MET antibody (10692-MM02), EGF antibody (0605-R008) were purchased from Sino Biological Inc. Recombinant human EGF (H6000–10), bFGF (H3000–10) were obtained from Beijing Wishbiotechnology Co., Ltd. Dynabeads MyOne streptavidin C1 magnetic beads (#65002) and Streptavidin Phycoerythrin Conjugates (S-866) were obtained from Invitrogen.

Techniques: Western Blot, Quantitation Assay

Journal: Cancers

Article Title: Lactate Oxidase Disrupts Lactate-Activated RAS and PI3K Oncogenic Signaling.

doi: 10.3390/cancers16162817

Figure Lengend Snippet: Figure 7. Model of lactate and LOX regulation of the HCAR1-RAS/PI3K signaling cascades. TME lactate activates HCAR1, which induces the assembly of a protein complex that includes the receptor itself, GRB2, SOS1, RAS, GAB1, p85, and p110β. Subsequently, the RAS and PI3K pathways are acti- vated and transduce the lactate-triggered signals, via phosphorylated ERK1/2 and AKT, respectively, to the downstream transcription factors that further mediate the transcription of the genes involved in promoting the signaling and cancer cell survival and growth (left panel). Introducing LOX to the TME depletes lactate and renders HCAR1 inactive for triggering the assembly of the RAS and PI3K signaling complex. The disassembled HCAR1-associated RAS and PI3K signaling proteins are targeted for degradation by proteasomes (right panel). TME, tumor microenvironment; PM, plasma membrane; TF, transcription factor; the criss cross lines in the right panel represent removal or loss of interaction or signaling.

Article Snippet: Antibodies for HCAR (Abcepta, CA, USA), GRB2 (Santa Cruz Biotechnology, TX, USA), SOS1 (Proteintech, IL, USA), KRAS (Santa Cruz Biotechnology, TX, USA), GAB1 (Novus Biologicals, CO, USA), p85 (Proteintech, IL, USA), p110β (Cell Signaling Technology, MA, USA), p110α (Boster Bio, CA, USA), pERK1/2 (Santa Cruz Biotechnology, TX, USA), ERK1/2 (Santa Cruz Biotechnology, TX, USA), AKT (Cell Signaling Technology, MA, USA), pAKTS473 (Cell Signaling Technology, MA, USA), pAKTT308 (Cell Signaling Technology, MA, USA), MCT1 (Santa Cruz Biotechnology, TX, USA), MCT4 (Santa Cruz Biotechnology, TX, USA), and Actin (Thermo Scientific, MA, USA) were used in this study.

Techniques: Clinical Proteomics, Membrane