Journal: Journal of Endocrinology

Article Title: Local adiponectin treatment reduces atherosclerotic plaque size in rabbits

doi: 10.1677/joe-06-0173

Figure Lengend Snippet: Figure 1 Serum adiponectin concentrations in rabbits. 1, Aorta non-injured rabbits on normal cholesterol diet; 2, aorta injured rabbits on high-cholesterol diet before adenovirus transfer; 3, after transfer Ad-bgal through intima; 4, after transfer Ad-APN through intima; 5, after transfer Ad-bgal through adventitia; 6, After transfer Ad-APN through adventitia. The symbol * indicates significant difference (P!0.01) vs serum adiponectin concentrations in aorta non-injured rabbits on normal cholesterol diet; † compared with rabbits before adenovirus transfer; and ‡ compared with rabbits after Ad-bgal transfer. The values are meansGS.E.M.

Article Snippet: Following blocking with 5% non-fat milk, the blots were incubated with anti-His tag mouse monoclonal antibody (1:1000 dilution, Applygen Technologies Inc., Beijing, China) and rabbit adiponectin polyclonal antibody (1:100 dilution, Boster, Wuhan, Hubei, China) at 4 8C overnight.

Techniques:

Journal: Journal of Endocrinology

Article Title: Local adiponectin treatment reduces atherosclerotic plaque size in rabbits

doi: 10.1677/joe-06-0173

Figure Lengend Snippet: Figure 2 SDS-PAGE (A) and western blot analysis of the supernatant of cultured Pichia pastoris (B, using anti-His tag monoclonal antibody; C, using adiponectin polyclonal antibody). Line 1 is a blank pGAPZa Pichia vector (not containing adiponectin gene) sample control and lines 2–4 are samples from the supernatant of cultured Pichia pastoris containing the recombinant rabbit adiponectin gene. Line 5 contained the molecular weight markers (not visible in image of blot). Both anti-His antibody and adiponectin polyclonal antibody detected a single predominant protein at z42 kDa (containing 9.3 kDa a-factor signal peptide and 2.5 kDa C-terminal His-tag).

Article Snippet: Following blocking with 5% non-fat milk, the blots were incubated with anti-His tag mouse monoclonal antibody (1:1000 dilution, Applygen Technologies Inc., Beijing, China) and rabbit adiponectin polyclonal antibody (1:100 dilution, Boster, Wuhan, Hubei, China) at 4 8C overnight.

Techniques: SDS Page, Western Blot, Cell Culture, Plasmid Preparation, Control, Recombinant, Molecular Weight

Journal: Journal of Endocrinology

Article Title: Local adiponectin treatment reduces atherosclerotic plaque size in rabbits

doi: 10.1677/joe-06-0173

Figure Lengend Snippet: Figure 5 Effect of Ad-APN treatment on adhesion molecules in atherosclerotic rabbits 14 days after recombinant adenovirus transfer. A, Immunohistochemical detection of adiponectin, VCAM-1 and ICAM-1 in atherosclerotic lesions. Representative images are shown. 1, Ad-bgal transfer through intima. 2, Ad-APN transfer through intima. 3, Ad-bgal transfer through adventitia. 4, Ad-APN transfer through adventitia. The magnification is 400. B, mRNA expression of VCAM-1 and ICAM-1 in abdominal aortic tissue in Ad-bgal or Ad-APN-treated rabbits. The values are meansGS.E.M.

Article Snippet: Following blocking with 5% non-fat milk, the blots were incubated with anti-His tag mouse monoclonal antibody (1:1000 dilution, Applygen Technologies Inc., Beijing, China) and rabbit adiponectin polyclonal antibody (1:100 dilution, Boster, Wuhan, Hubei, China) at 4 8C overnight.

Techniques: Recombinant, Immunohistochemical staining, Expressing

Journal: Acta Biochimica et Biophysica Sinica

Article Title: Diacylglycerol kinase γ facilitates the proliferation and migration of neural stem cells in the developing neural tube

doi: 10.3724/abbs.2024156

Figure Lengend Snippet: Immunofluorescence staining showing the coexpression of DGKγ with p-PKCγ or p-PKCδ DGKγ was coexpressed with p-PKCγ (A) and p-PKCδ (B) in the cytoplasm and nucleus of neuroepithelial NT cells at E11.5. Blue: DAPI; red: DGKγ; green: p-PKCγ or p-PKCδ. Scale bar: 100 μm. n = 5.

Article Snippet: The PVDF membranes were subsequently incubated with primary antibodies against DGKγ (1:500, ab89037; Abcam, Cambridge, UK), p-PKCδ (1:1000, sc-365969; Santa Cruz Biotech), PKCδ (1:1000, sc-213; Santa Cruz Biotech), p-PKCγ (1:1000, 44-975G; Invitrogen), PKCγ (1:1000, BM5623; Boster), and β-actin (1:4000, AP0060; Bioworld Technology, Shanghai, China) overnight at 4°C.

Techniques: Immunofluorescence, Staining

Journal: Acta Biochimica et Biophysica Sinica

Article Title: Diacylglycerol kinase γ facilitates the proliferation and migration of neural stem cells in the developing neural tube

doi: 10.3724/abbs.2024156

Figure Lengend Snippet: Changes in DAG content and p-PKC subtypes during NSC migration (A) DAG contents were determined by ELISA, which revealed that the DAG contents were significantly increased in sh-DGKγ-treated cells compared with those in the sh-NC-treated cells and significantly increased in R59949- and PMA-treated cells compared with those in the vehicle-treated cells. (B) Representative western blots of p-PKCγ and p-PKCδ in NSCs were quantified via western blotting. (C,D) Densitometric analysis of the western blots revealed that the p-PKCγ/PKCγ ratio was significantly lower in the PMA group than in the vehicle group; however, the p-PKCδ/PKCδ ratio was significantly greater in the sh-DGKγ group than in the sh-NC group, which was also significantly greater in the R59949 and PMA groups than in the vehicle group. *P < 0.05, **P < 0.01, ***P < 0.001 vs sh-NC or vehicle. n = 3.

Article Snippet: The PVDF membranes were subsequently incubated with primary antibodies against DGKγ (1:500, ab89037; Abcam, Cambridge, UK), p-PKCδ (1:1000, sc-365969; Santa Cruz Biotech), PKCδ (1:1000, sc-213; Santa Cruz Biotech), p-PKCγ (1:1000, 44-975G; Invitrogen), PKCγ (1:1000, BM5623; Boster), and β-actin (1:4000, AP0060; Bioworld Technology, Shanghai, China) overnight at 4°C.

Techniques: Migration, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Acta Biochimica et Biophysica Sinica

Article Title: Diacylglycerol kinase γ facilitates the proliferation and migration of neural stem cells in the developing neural tube

doi: 10.3724/abbs.2024156

Figure Lengend Snippet: Schematic diagram of the role of DGKγ in NSC migration and proliferation A schematic diagram showing that the DAG/PKCδ signaling pathway is involved in NSC migration and that the mTOR signaling pathway, independent of PKC, may be involved in NSC proliferation.

Article Snippet: The PVDF membranes were subsequently incubated with primary antibodies against DGKγ (1:500, ab89037; Abcam, Cambridge, UK), p-PKCδ (1:1000, sc-365969; Santa Cruz Biotech), PKCδ (1:1000, sc-213; Santa Cruz Biotech), p-PKCγ (1:1000, 44-975G; Invitrogen), PKCγ (1:1000, BM5623; Boster), and β-actin (1:4000, AP0060; Bioworld Technology, Shanghai, China) overnight at 4°C.

Techniques: Migration

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Autologous fat transplants to deliver glitazone and adiponectin for vasculoprotection

doi: 10.1016/j.jconrel.2017.08.036

Figure Lengend Snippet: Shown is the concentration of adiponectin detected in the media removed at day 4. N = 5 separate experiments using fat from five subjects. *P<0.05, 0 μM PGZ vs 60 μM PGZ; **P<0.01, 0 μM PGZ vs 600 μM PGZ.

Article Snippet: Nonspecific binding was blocked with 2% goat serum and tissue slices were incubated overnight at 4 °C with a rabbit anti-porcine adiponectin antibody (1:125 dilution) (Xeno Diagnostics, LLC).

Techniques: Concentration Assay

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Autologous fat transplants to deliver glitazone and adiponectin for vasculoprotection

doi: 10.1016/j.jconrel.2017.08.036

Figure Lengend Snippet: Fat with or without PGZ (200 μM) was placed perivascular to the left and right external jugular vein, respectively, in a pig. The veins were explanted at 2 weeks post-transplantation. The tissue was immunostained with anti-adiponectin antibody. Images A and B are from the vein exposed to the fat transplant with PGZ. Images C and D are from the vein exposed to the fat transplant without PGZ. Images A and B are of the vein wall and B and D are of the adipose tissue perivascular to the vessel wall. Dark gray indicates positive antibody binding. The length bar indicates 200 μm.

Article Snippet: Nonspecific binding was blocked with 2% goat serum and tissue slices were incubated overnight at 4 °C with a rabbit anti-porcine adiponectin antibody (1:125 dilution) (Xeno Diagnostics, LLC).

Techniques: Transplantation Assay, Binding Assay