Journal: EBioMedicine

Article Title: Proapoptotic Cyclic Peptide BC71 Targets Cell-Surface GRP78 and Functions as an Anticancer Therapeutic in Mice

doi: 10.1016/j.ebiom.2018.06.004

Figure Lengend Snippet: BC71 targets cell-surface GRP78 but not αvβ5 integrin to induce apoptosis. (a) BC71 induces HUVECs apoptosis in a dose-dependent manner. HUVECs were treated with BC71 (concentration range: 12.5, 25, 50, 100 μM) for 24 h and apoptosis was determined using the cell death ELISA kit (Roche). (b) Anti-GRP78 N-terminal domain antibody blocked the apoptosis function of BC71 in a dose-dependent manner. The apoptosis of the combined treatment with increasing amount of anti-GRP78 N-terminal domain antibody and 100 μM BC71 for 24 h was measured using the Cell Death Detection ELISA. (c) Anti-GRP78 C-terminal domain antibody and (d) anti-αvβ5 antibody did not block BC71 induced apoptosis. For clarity of presentation, data were normalized with that of non-treated (VEGF only) cells, which was set as 1. Data are expressed as mean ± standard error of the mean. The results are representative of at least three independent experiments. Statistical significance was determined using ANOVA. *P < 0.05; **P < 0.01, n ≥ 3.

Article Snippet: Antibodies against integrin αvβ5 (P1F76; Santa Cruz Biotechnology, Santa Cruz, CA, USA), GRP78 (A-10, Santa Cruz Biotechnology) and GRP78 (C-20, Santa Cruz Biotechnology) were used for neutralization.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Blocking Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Acupuncture Delays Cartilage Degeneration through Upregulating SIRT1 Expression in Rats with Osteoarthritis

doi: 10.1155/2021/2470182

Figure Lengend Snippet: Acupuncture inhibited OA-associated inflammation, the NF- κ B signaling pathway, and ECM degradation through upregulating SIRT1 expression in rat articular cartilages. (a/b): The levels of TNF- α and IL-2 in the serum of acupuncture-treated OA rats injected with or without shSIRT1 lentiviruses were assessed by enzyme-linked immunosorbent assay. (c/d/e/f/g/h): The expressions of SIRT1, MMP-9, ADAMTS5, p-p65/p65, p-I κ B α /I κ B α , collagen II, and aggrecan in the articular cartilage of acupuncture-treated OA rats injected with or without shSIRT1 lentiviruses were analyzed by western blot, with GAPHD serving as a reference gene. ∧ P or ‡ P < 0.05; ∗∗ P or ∧∧ P or ## P or ‡‡ P < 0.01; ∗∗∗ P or ^^^ P or ### P or ‡‡‡ P < 0.001; ∗ vs. Sham; ∧ vs. Model + shNC; # vs. Model + Acupuncture + shNC; ‡ vs. Model + shSIRT1 (OA: Osteoarthritis; TNF- α : Tumor necrosis factor- α ; IL-2: interleukin-2; shNC: shRNA-negative control; MMP-9: matrix metallopeptidase-9; SIRT1: NAD-dependent deacetylase sirtuin-1; ADAMTS5: a disintegrin and metalloproteinase with thrombospondin motifs 5).

Article Snippet: The membranes were blocked by 5% nonfat milk (P2194, Sigma-Aldrich, USA) in tris buffered saline with 1% Tween 20 (TBST; TA-125-TT, ThermoFisher, USA) for 1 h and further probed with primary antibodies against SIRT1 (#9475, 120 kDa, 1 : 1000, Cell Signaling Technology, Danvers, MA, USA), matrix metallopeptidase (MMP)-9 (ab76003, 92 kDa, 1 : 1000, Abcam, USA), A disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5; ab41037, 73 kDa, 1 : 250, Abcam, USA), phosphorylated (p)-p65 (#3033, 62 kDa, 1 : 1000, Cell Signaling Technology, USA), p65 (#8242, 65 kDa, 1 : 1000, Cell Signaling Technology, USA), p-I κ B α (#2859, 40 kDa, 1 : 1000, Cell Signaling Technology, USA), I κ B α (#4812, 39 kDa, 1 : 1000, Cell Signaling Technology, USA), collagen II (ab188570, 141 kDa, 1 : 1000, Abcam, USA), aggrecan (orb624552, 250 kDa, 1 : 500, Biorbyt, Cambridge, UK), and GAPDH (ab8245, 36 kDa, 1 : 1000, Abcam, USA) at 4°C overnight.

Techniques: Expressing, Injection, Enzyme-linked Immunosorbent Assay, Western Blot, shRNA, Negative Control, Histone Deacetylase Assay

Journal: Breast Cancer Research

Article Title: Tumor-specific expression of αvβ3 integrin promotes spontaneous metastasis of breast cancer to bone

doi: 10.1186/bcr1398

Figure Lengend Snippet: αvβ3 integrin expression in tumor lines of the metastasis model. Cell surface expression of integrin subunits was determined by measuring the specific fluorescence by flow cytometry as described in Materials and methods. Shaded area, isotype; dotted line, αv; solid line, β3.

Article Snippet: The cells were then incubated with 2 μg/ml hamster anti-mouse-αv, anti-β3 or isotype control primary antibody (BD Pharmingen, North Ryde, NSW, Australia) diluted in labeling buffer (α-MEM supplemented with 2% FCS) for 1 hour on ice.

Techniques: Expressing, Fluorescence, Flow Cytometry