Journal: Cells

Article Title: Caspase-Dependent HMGB1 Release from Macrophages Participates in Peripheral Neuropathy Caused by Bortezomib, a Proteasome-Inhibiting Chemotherapeutic Agent, in Mice

doi: 10.3390/cells10102550

Figure Lengend Snippet: Involvement of HMGB1-targeted receptors in the CIPN caused by bortezomib. Bortezomib at 0.4 mg/kg or vehicle was administered i.p. on days 0, 2, 5, 7, 9, and 12. ( A , B ) Preventive ( A ) or therapeutic ( B ) effect of FPS-ZM1, TAK-242, and AMD3100, antagonists of RAGE, TLR4, and CXCR4, respectively. The mice received repeated i.p. administration of FPS-ZM1 at 1 mg/kg, TAK-242 at 3 mg/kg, AMD3100 at 8 mg/kg, or vehicle, 30 min before each dose of bortezomib ( A ), or single i.p. administration of each of them on day 14 ( B ). ( C , D ) Protein levels of RAGE, TLR4, and CXCR4 in the dorsal root ganglion ( C ) and sciatic nerves ( D ) on day 14 after the onset of bortezomib treatment. ( E , F ) Lack of preventive ( E ) and therapeutic ( F ) effects of TH1020, a TLR5 antagonist. The mice received repeated ( E ) or single ( F ) i.p. administration of TH1020 at 1 mg/kg or vehicle, in mice treated with bortezomib according to the above-mentioned schedules. Data show the mean with S.E.M for 5–6 ( A , B , F ), 5–7 ( C ), 6–7 ( D ), or 6 ( E ) mice. V, vehicle; BTZ, bortezomib; DRG, dorsal root ganglion; * p < 0.05, ** p < 0.01 vs. V ( C ) or V in V-treated mice ( A , B , E , F ). † p < 0.05, †† p < 0.01 vs. V in BTZ-treated mice.

Article Snippet: Primary antibodies were: an anti-HMGB1 rabbit polyclonal antibody (Abcam, Cambridge, UK) (1: 5000 dilution), anti-RAGE rabbit polyclonal antibody (Abcam) (1:1000 dilution), anti-TLR4 rabbit polyclonal antibody (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) (1:10000 dilution), anti-CXCR4 rabbit polyclonal antibody (Novus Biologicals, Littleton, CO, USA) (1:1000 dilution), and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) rabbit polyclonal antibody (Santa Cruz Biotechnology, Inc.) (1:5000 dilution).

Techniques:

Journal: Cells

Article Title: Caspase-Dependent HMGB1 Release from Macrophages Participates in Peripheral Neuropathy Caused by Bortezomib, a Proteasome-Inhibiting Chemotherapeutic Agent, in Mice

doi: 10.3390/cells10102550

Figure Lengend Snippet: Scheme for bortezomib-induced caspase-dependent HMGB1 release from macrophages and CIPN development, in contrast to caspase-independent mechanisms for paclitaxel. ( A ) Inhibition of proteasome by bortezomib causes caspase-dependent apoptosis of macrophages followed by the release of HMGB1, which in turn causes neuronal sensitization via activation of RAGE and acceleration of CXCL12/CXCR4 signals, leading to CIPN. ( B ) Paclitaxel causes HMGB1 release from macrophages through activation of the ROS/p38MAPK/NF-κB pathway , independently of caspase (see D), and the extracellular HMGB1 develops CIPN in a manner dependent on RAGE and CXCR4 , as shown in the CIPN caused by bortezomib.

Article Snippet: Primary antibodies were: an anti-HMGB1 rabbit polyclonal antibody (Abcam, Cambridge, UK) (1: 5000 dilution), anti-RAGE rabbit polyclonal antibody (Abcam) (1:1000 dilution), anti-TLR4 rabbit polyclonal antibody (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) (1:10000 dilution), anti-CXCR4 rabbit polyclonal antibody (Novus Biologicals, Littleton, CO, USA) (1:1000 dilution), and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) rabbit polyclonal antibody (Santa Cruz Biotechnology, Inc.) (1:5000 dilution).

Techniques: Inhibition, Activation Assay

Journal: The European respiratory journal

Article Title: Transforming growth factor-beta1 in sarcoidosis.

doi: 10.1183/09031936.98.12040913

Figure Lengend Snippet: Fig. 1. – Immunohistochemical localization of transforming growth factor (TGF)-β1 in lung tissue. Lung biopsies obtained from patients with sar- coidosis were examined to determine the localization of TGF-β1. A-D) representative sections of lung sections from patients with sarcoidosis: A) TGF- β1 was detected predominantly within the non-necrotizing granuloma; B) TGF-β1 was localized predominantly in the epithelioid histiocytes comprising non-necrotizing granuloma; C) bronchiolar epithelium was positive for TGF-β1 expression; D) TGF-β1 staining was detected in alveolar macrophages (AM) and hyperplastic type II pneumocytes. E) representative section of lung section from an untreated patient with idiopathic pulmonary fibrosis. TGF-β1 antibody is seen in alveolar epithelial cells lining cystic spaces and in AM, and is also distributed in association with fibrous connective tissue bordering the epithelial cells. F) representative section of lung section from a control. TGF-β1 was localized in some AM. (Internal scale bar: A=50 µm, B–F=25 µm.)

Article Snippet: The antibody used for immunohistochemical staining was a commercially available mouse monoclonal antibody directed against TGF-β1 (epitope corresponding to amino acids 328–353 mapping at the C-terminus of the precursor form of human TGF-β1; Santa-Cruz Biotechnology, CA, USA).

Techniques: Immunohistochemical staining, Expressing, Staining, Control

Journal: Scientific reports

Article Title: Soluble VEGF receptor 1 (sFLT1) induces non-apoptotic death in ovarian and colorectal cancer cells.

doi: 10.1038/srep24853

Figure Lengend Snippet: Figure 1. Soluble FLT1 has a suppressive effect against cell proliferation, and the effect is neutralized by VEGF. (a) The number of cells transfected with pLV-EGFP or pLV-sFLT1 as well as with addition of VEGF. The number of cells was significantly lower in the sFLT1 group. *P < 0.05 compared to the pLV-EGFP group. (b) The number of cells transfected by pLV-EGFP as well as addition of rVEGFR1. In SKOV3 and HeyA8 cell lines, the cell numbers were significantly lower in the recombinant VEGFR1 groups compared to the control groups. *P < 0.05 compared to the pLV-EGFP group. P values are as follows; *1:P = 0.024, *2:P = 0.025, *3:P = 0.018, *4:P = 6.8 × 10−4, *5:P = 0.022, *6:P = 2.1 × 10−3, *7:P = 5.3 × 10−3, *8:P = 0.030, *9:P = 0.039, *10:P = 0.041, *11:P = 0.033, *12:P = 0.013.

Article Snippet: Recombinant Human VEGF 165 (rVEGF) and recombinant Human VEGF R1 (rVEGFR1) (27–328 amino acids corresponding to the extracellular region of cell-surface FLT1) was purchased from R&D Systems.

Techniques: Transfection, Recombinant, Control

Journal: Scientific reports

Article Title: Soluble VEGF receptor 1 (sFLT1) induces non-apoptotic death in ovarian and colorectal cancer cells.

doi: 10.1038/srep24853

Figure Lengend Snippet: Figure 3. Both transfected and exogenously applied sFLT1/rVEGFR1 induce necrosis. (a) Morphology of HEK293T, SKOV3, HT-29, and HeyA8 cells, which were visualized under phase-contrast optics. 1: No treatment. 2: HEK293T, SKOV3, HT-29, and HeyA8 cells transfected pLV-EGFP. 3: HEK293T, SKOV3, HT-29, and HeyA8 cells transfected with pLV-sFLT1 were larger, and cell adhesion was disturbed. 4: HEK293T, SKOV3, HT-29, and HeyA8 cells treated with 2.5 mM H2O2 to induce necrosis. The rupture of the cell membrane and swelling of the cells were observed. 5: Treatment of HEK293T, SKOV3, HT-29, and HeyA8 cells treated with

Article Snippet: Recombinant Human VEGF 165 (rVEGF) and recombinant Human VEGF R1 (rVEGFR1) (27–328 amino acids corresponding to the extracellular region of cell-surface FLT1) was purchased from R&D Systems.

Techniques: Transfection, Membrane

Journal: Scientific reports

Article Title: Soluble VEGF receptor 1 (sFLT1) induces non-apoptotic death in ovarian and colorectal cancer cells.

doi: 10.1038/srep24853

Figure Lengend Snippet: Figure 5. Soluble FLT1 shows an anti-tumour effect in mice injected with SKOV3 cells. Female nude mice were inoculated subcutaneously with SKOV3 cells. Treatments started when tumour volume exceeded 10 mm3, as described in Materials and Methods. (a) Anti-tumour activity of intraperitoneal administration of rVEGFR1. Recombinant VEGFR1-2,000 ng, rVEGFR1–200 ng and bevacizumab groups exhibited significantly smaller tumours compared to the PBS group (P = 0.0012, 0.0097, and 0.0038, respectively). Data points indicate the mean values ± S.E. of the tumour volume. Statistically significant differences are indicated by asterisks: *P < 0.05 significantly different from PBS treated mice. Body weight loss was not observed after treatment with rVEGFR1 or bevacizumab. (b) Anti-tumour activity of transfected sFLT1 in SKOV3 cells. The pLV-sFLT1-transfected group exhibited significantly smaller tumours compared to the pLV-EGFP-transfected group (P = 8.2 × 10−4). Data points indicate the mean values ± S.E. of tumour volume. Statistically significant differences are indicated by asterisks: **P < 0.05 significantly different from pLV-EGFP mice. (c) Representative images of apoptotic nuclear TUNEL staining using a Dead End™ fluorometric TUNEL assay kit. TUNEL- positive cells were visualized with green fluorescent protein at 20× magnification. Propidium iodide stains nucleus. TUNEL-positive staining was rarely observed in cells either exogenously rVEGFR1 or bevacizumab treated tumours. (d) Effect of the sFLT1 treatment on body weight. Each bar represents the mean ± S.E.

Article Snippet: Recombinant Human VEGF 165 (rVEGF) and recombinant Human VEGF R1 (rVEGFR1) (27–328 amino acids corresponding to the extracellular region of cell-surface FLT1) was purchased from R&D Systems.

Techniques: Injection, Activity Assay, Recombinant, Transfection, TUNEL Assay, Staining

Journal: Endocrinology

Article Title: Muscle-Specific Deletion of Comparative Gene Identification-58 (CGI-58) Causes Muscle Steatosis but Improves Insulin Sensitivity in Male Mice

doi: 10.1210/en.2014-1892

Figure Lengend Snippet: Reduced TG hydrolase activity and fatty acid oxidation in the heart of mCgi58-KO mice. A, Cardiac TG hydrolase activity (n = 5). B, Western blottings of ATGL. GAPDH was used as a loading control. C, Relative mRNA levels in the ventricular tissue of control and mCgi58-KO mice (n = 4). D, Cardiac fatty acid oxidation activity (n = 5). E, Cardiac contents of total acyl-carnitine (AC) and AC species (n = 6). All tissue samples were collected from 4-hour-fasted mice fed the HFD for 12 weeks. *, P < .05; **, P < .01.

Article Snippet: A rabbit polyclonal antibody against human ATGL amino acids 328–400 was obtained from Cayman Chemical (1006409) and this antibody cross-reacts with human, mouse, and rat ATGL.

Techniques: Activity Assay, Western Blot, Control