Journal: Arthritis Research & Therapy

Article Title: Expression of ADAM15 in rheumatoid synovium: up-regulation by vascular endothelial growth factor and possible implications for angiogenesis

doi: 10.1186/ar1796

Figure Lengend Snippet: Effects of passages, starvation, cytokines and growth factors on ADAM15 expression in cultured cells. The mRNA expression of ADAM15 was examined by RT-PCR at 25 cycles as described in Materials and methods. (a) Effect of passages on the mRNA expression of ADAM15 in rheumatoid arthritis (RA) synovial fibroblasts (SFs). Lanes 1 to 5 indicate passages 5, 6, 7, 8 and 9 of RA SFs. (b) Effect of starvation on the mRNA expression of ADAM15 in RA SFs. (c) Effect of tumor necrosis factor (TNF)-α (0, 0.1, 1 and 10 ng/ml), IL-1α (0, 0.1, 1 and 10 ng/ml) or transforming growth factor (TGF)-β (0, 0.1, 1 and 10 ng/ml) on the mRNA expression of ADAM15 in RA SFs after stimulation with these factors for 24 h. (d) Regulation of the mRNA expression of ADAM15 by vascular endothelial growth factor (VEGF) 165 in RA SFs and human umbilical vein endothelial cells (HUVECs). Cells were stimulated with VEGF 165 (0, 1, 10 and 50 ng/ml) for 24 h. Note that VEGF 165 enhances the expression of ADAM15 only in HUVECs.

Article Snippet: To exclude the possible involvement of VEGFR-1 in ADAM15 expression, RA SFs and HUVECs were stimulated with recombinant human placenta growth factor (PlGF; 1, 10 or 50 ng/ml; R&D Systems), which selectively binds to VEGFR-1 [ ].

Techniques: Expressing, Cell Culture, Reverse Transcription Polymerase Chain Reaction

Journal: Arthritis Research & Therapy

Article Title: Expression of ADAM15 in rheumatoid synovium: up-regulation by vascular endothelial growth factor and possible implications for angiogenesis

doi: 10.1186/ar1796

Figure Lengend Snippet: Effects of cytokines and growth factors on expression of vascular endothelial growth factor receptors (VEGFRs). The mRNA expression of the VEGFRs was examined by RT-PCR at 30 cycles as described in Materials and methods. (a) The expression of VEGFR-1, VEGFR-2 and neuropilin-1 in rheumatoid arthritis (RA) synovial fibroblasts (SFs) of different passages and human umbilical vein endothelial cells (HUVECs). Lanes 1 to 5 correspond to RA SFs of passages 5, 6, 7, 8 and 9, respectively. (b) The mRNA expression of VEGFR-1, VEGFR-2 and neuropilin-1 in RA SFs after 24 h stimulation with tumor necrosis factor (TNF)-α (0, 0.1, 1, 10 and 50 ng/ml), IL-1α (0, 0.1, 1, 10 and 50 ng/ml) or transforming growth factor (TGF)-β (0, 0.01, 0.1, 1 and 10 ng/ml). Note that TNF-α selectively induces the mRNA expression of VEGFR-2 in RA SFs.

Article Snippet: To exclude the possible involvement of VEGFR-1 in ADAM15 expression, RA SFs and HUVECs were stimulated with recombinant human placenta growth factor (PlGF; 1, 10 or 50 ng/ml; R&D Systems), which selectively binds to VEGFR-1 [ ].

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Arthritis Research & Therapy

Article Title: Expression of ADAM15 in rheumatoid synovium: up-regulation by vascular endothelial growth factor and possible implications for angiogenesis

doi: 10.1186/ar1796

Figure Lengend Snippet: Immunohistochemistry of ADAM15, vascular endothelial growth factor receptor (VEGFR)-2 and von Willebrand factor (vWF). (a-d) Rheumatoid arthritis (RA) synovial fibroblasts (SFs) and (e-h) human umbilical vein endothelial cells (HUVEC) were cultured on Lab-Tek II chamber slides and immunostained with antibodies against (a,e) ADAM15, (b,f) VEGFR-2 or (c,g) vWF or (d,h) non-immune mouse IgG as described in Materials and methods. Immunostaining of VEGFR-2 in RA SFs (b) was performed with RA SFs that were treated with 10 ng/ml TNF-α for 24 h prior to immunohistochemistry. Note that vWF is not immunostained in RA SFs (c) , but VEGFR-2 is expressed in those stimulated with TNF-α (b) . Scale bar, 25 μm.

Article Snippet: To exclude the possible involvement of VEGFR-1 in ADAM15 expression, RA SFs and HUVECs were stimulated with recombinant human placenta growth factor (PlGF; 1, 10 or 50 ng/ml; R&D Systems), which selectively binds to VEGFR-1 [ ].

Techniques: Immunohistochemistry, Cell Culture, Immunostaining

Journal: Journal of Virology

Article Title: Palmitoylation enhances the stability of porcine epidemic diarrhea virus spike protein by antagonizing its degradation via chaperone-mediated autophagy to facilitate viral proliferation

doi: 10.1128/jvi.00347-25

Figure Lengend Snippet: Palmitoylation of PEDV S protein enhances its stability. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The cells were treated with or without 20 µM 2-BP for 36 h. The mRNA abundance of S protein was quantified by RT-qPCR. Statistical analysis was carried out using Student’s t -test. ns, not significant ( P > 0.05). ( B ) Vero cells were transfected with the plasmid encoding PEDV S-Flag and were then treated with 2-BP (5 µM, 10 µM, and 20 µM) or DMSO for 36 h. The S protein levels were measured by IB. ( C ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector for 36 h. The S protein levels were measured by IB. ( D ) Vero cells were transfected with si- ZDHHC5 or si-NC for 12 h and were then transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector for 24 h. The S protein levels were measured by IB. ( E ) Vero cells were co-transfected with the plasmids encoding S-Flag and ZDHHC5-Myc (0.5 µg, 1 µg, and 2 µg) for 36 h. The S protein levels were measured by IB. ( F ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. The S-Flag-overexpressed cells were treated with or without 20 µM 2-BP for 24 h. Then, the cells were stimulated with CHX (1 µg/mL) and lysed at the indicated time points (0 h, 2 h, 4 h, 6 h, and 8 h). The samples were analyzed by IB. ( G ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or Flag-tagged empty vector. The cells were treated with DMSO or 20 µM 2-BP, and then co-incubated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( H ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. The cells were treated with DMSO, 3-MA (2 mM), carfilzomib (20 nM), NH 4 Cl (0.5 mM), or CQ (40 µM) for 36 h. The cells were lysed and analyzed by IB. ( I ) Vero cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. At 6 h post-transfection, the S-Flag-overexpressed groups were treated with or without 20 µM 2-BP, and all the cells were treated with NH 4 Cl (0.5 mM) for 24 h. The lysosomes were isolated from these cells and analyzed by IB. The mean gray values of S protein were quantified using ImageJ software.

Article Snippet: TransIntro PL transfection reagent (Cat. No. FT301-01) was purchased from TransGen Biotech.

Techniques: Transfection, Plasmid Preparation, Quantitative RT-PCR, Incubation, Isolation, Software

Journal: Journal of Virology

Article Title: Palmitoylation enhances the stability of porcine epidemic diarrhea virus spike protein by antagonizing its degradation via chaperone-mediated autophagy to facilitate viral proliferation

doi: 10.1128/jvi.00347-25

Figure Lengend Snippet: The KFERQ-like motif (QVDRL) in PEDV S protein is recognized and bound by HSC70. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-ΔCRR or Flag-tagged empty vector for 36 h. The proteins were immunoprecipitated in WCL using anti-Flag magnetic beads, and then the associated proteins were separated by 7.5% SDS-PAGE and stained with silver. The black arrow indicated the expressed S-ΔCRR, and the red arrow indicated a different immunoprecipitated protein band in the S-ΔCRR-expressed cells. Lane M, protein marker. ( B ) The silver-stained protein band indicated by the red arrow was subjected to LC-MS/MS. ( C ) HEK-293T cells were co-transfected with the plasmids encoding PEDV S-Flag and HSC70-Myc, with Flag-tagged or Myc-tagged empty vector as control for 36 h. The supernatant of WCL was immunoprecipitated using anti-Flag magnetic beads or anti-Myc magnetic beads. The precipitated proteins were analyzed by IB. ( D ) Vero cells were co-transfected with the plasmids encoding PEDV S-Flag and HSC70-Myc for 24 h. The cells were visualized with the specific primary and secondary antibodies. Cell nuclei were stained with DAPI. Images were taken at a 630× magnification and representative of a single slice of a stack from three independent experiments. Representative images are shown. Scale bars, 10 µm. The co-localization was assessed by determination of the Pearson’s correlation coefficient using the JaCoP plugin in ImageJ software. The mean value ± SEM is representative of three individual enlarged pictures. ( E ) Analyses of KFERQ-like motifs in PEDV S proteins. The KFERQ-like motif typically consists of constant glutamine (Q) flanked on one side of the pentapeptide sequence, followed by one or two positively charged amino residues (lysine, [K] or arginine, [R]), one or two hydrophobic amino residues (phenylalanine, [F]; isoleucine, [I]; leucine, [L]; or valine, [V]), and one negatively charged amino residue (aspartate, [D] and glutamic acid, [E]) . ( F ) HEK-293T cells were co-transfected with the plasmids encoding HSC70-Myc and PEDV S-Flag, S Q1082A , S Q1117A , S Q1082A/Q1117A , or Flag-tagged empty vector. At 36 h post-transfection, the supernatant of WCL was immunoprecipitated using anti-Flag magnetic beads. The precipitated proteins were analyzed by IB.

Article Snippet: TransIntro PL transfection reagent (Cat. No. FT301-01) was purchased from TransGen Biotech.

Techniques: Transfection, Plasmid Preparation, Immunoprecipitation, Magnetic Beads, SDS Page, Staining, Marker, Liquid Chromatography with Mass Spectroscopy, Control, Software, Sequencing, Residue

Journal: Journal of Virology

Article Title: Palmitoylation enhances the stability of porcine epidemic diarrhea virus spike protein by antagonizing its degradation via chaperone-mediated autophagy to facilitate viral proliferation

doi: 10.1128/jvi.00347-25

Figure Lengend Snippet: Palmitoylation of PEDV S protein prevents its QVDRL motif from being recognized by HSC70. ( A ) HEK-293T cells were transfected with the plasmids encoding HSC70-Myc and PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector. At 6 h post-transfection, the S-Flag-overexpressed cells were treated with or without 40 µM 2-BP. After 36 h post-transfection, the supernatant of WCL was immunoprecipitated using anti-Flag magnetic beads, and the precipitated proteins were analyzed by IB. ( B ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. At 6 h post-transfection, the S-Flag-overexpressed groups were treated with or without 20 µM 2-BP. At 24 h post-transfection, S-Flag, S-ΔCRR, or HSC70 was visualized with the specific primary and secondary antibodies. Cell nuclei were stained with DAPI. Images were taken at a 630× magnification and representative of a single slice of a stack from three independent experiments. Representative images are shown. Scale bars, 10 µm. The co-localization was assessed by determination of the Pearson’s correlation coefficient using the JaCoP plugin in ImageJ software. The mean value ± SEM is representative of three individual enlarged pictures. Statistical analysis was carried out using Student’s t -test. ** P < 0.01. ( C ) HEK-293T cells were co-transfected with the plasmids encoding HSC70-Myc and PEDV S-Flag, S-ΔCRR, S Q1082A , S-ΔCRR Q1082A , or Flag-tagged empty vector. At 6 h post-transfection, the S-Flag-overexpressed and the S Q1082A -overexpressed cells were treated with or without 40 µM 2-BP. Subsequent assays were performed as described for panel A. (D through F) Vero cells were transfected with si- HSC70 or si-NC. At 12 h post-transfection, the cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector, and treated with or without 20 µM 2-BP for another 24 h. The cells were lysed and analyzed by IB. The mean gray values of S protein and HSC70-Myc were quantified using ImageJ software.

Article Snippet: TransIntro PL transfection reagent (Cat. No. FT301-01) was purchased from TransGen Biotech.

Techniques: Transfection, Plasmid Preparation, Immunoprecipitation, Magnetic Beads, Staining, Software

Journal: Journal of Virology

Article Title: Palmitoylation enhances the stability of porcine epidemic diarrhea virus spike protein by antagonizing its degradation via chaperone-mediated autophagy to facilitate viral proliferation

doi: 10.1128/jvi.00347-25

Figure Lengend Snippet: Palmitoylation of PEDV S protein prevents its degradation via CMA. ( A ) Vero cells were transfected with the plasmid encoding PEDV S-Flag or S-ΔCRR. At 6 h post-transfection, the S-Flag-overexpressed cells were treated with or without 20 µM 2-BP and then treated with NH 4 Cl (0.5 mM). At 24 h post-transfection, S-Flag, S-ΔCRR, or LAMP2A was visualized with the specific primary and secondary antibodies. Cell nuclei were stained with DAPI. Images were taken at a 630× magnification and representative of a single slice of a stack from three independent experiments. Representative images are shown. Scale bars, 10 µm. The co-localization was assessed by determination of the Manders’ overlap coefficient using the JaCoP plugin in ImageJ software. The mean value ± SEM is representative of three individual enlarged pictures. Statistical analysis was carried out using Student’s t -test. ** P < 0.01; *** P < 0.001. (B through D) Vero cells were transfected with si- LAMP2A or si-NC. At 12 h post-transfection, the cells were transfected with the plasmid encoding PEDV S-Flag, S-ΔCRR, or Flag-tagged empty vector and treated with or without 20 µM 2-BP for another 24 h. The cells were lysed and analyzed by IB. The mean gray values of S protein were quantified using ImageJ software.

Article Snippet: TransIntro PL transfection reagent (Cat. No. FT301-01) was purchased from TransGen Biotech.

Techniques: Transfection, Plasmid Preparation, Staining, Software

Journal: Oncotarget

Article Title: Pleiotrophin enhances PDGFB-induced gliomagenesis through increased proliferation of neural progenitor cells

doi: 10.18632/oncotarget.12983

Figure Lengend Snippet: A. qPCR and western blot analysis of tv-a expressing NPCs treated with virus-containing media from DF-1 cells transfected with RCAS-ev or RCAS-PTN. B. Glioma incidence in Gtv-a;Arf mice infected with RCAS-ev (empty vector, negative control), RCAS-PTN or RCAS-PDGFB (positive control) virus. C. Glioma incidence in G/tv-a wild type mice infected with RCAS-ev in combination with RCAS-PDGFB or RCAS-PTN in combination with RCAS-PDGFB.

Article Snippet: After 2 hours, the cells were treated with 25ng/ml PTN (252-PL-050, R&D systems), 10ng/ml PDGFB (100-14b, Peprotech) or the combination of PTN and PDGFB.

Techniques: Western Blot, Expressing, Virus, Transfection, Infection, Plasmid Preparation, Negative Control, Positive Control

Journal: Oncotarget

Article Title: Pleiotrophin enhances PDGFB-induced gliomagenesis through increased proliferation of neural progenitor cells

doi: 10.18632/oncotarget.12983

Figure Lengend Snippet: A. H&E stain and immunohistochemical staining of MAP2 and GFAP. B. Distribution of pathological grades. Results were plotted as percentage of samples per grade in G/tv-a wt mice injected with the RCAS-PDGFB+RCAS-ev ( n = 12) or RCAS-PDGFB+ RCAS-PTN ( n = 22). (GII: grade II, GIII: grade III). C. - E. Immunohistochemical staining of CD31 and stereological quantification of vessel area and vessel diameter in RCAS-PDGFB+RCAS-ev and RCAS-PDGFB+RCAS-PTN grade II and grade III gliomas (means ± SD; n = 3-5 mice/group). (Bar = 20 μm in A, Bar = 100 μm in C)

Article Snippet: After 2 hours, the cells were treated with 25ng/ml PTN (252-PL-050, R&D systems), 10ng/ml PDGFB (100-14b, Peprotech) or the combination of PTN and PDGFB.

Techniques: Staining, Immunohistochemical staining, Injection

Journal: Oncotarget

Article Title: Pleiotrophin enhances PDGFB-induced gliomagenesis through increased proliferation of neural progenitor cells

doi: 10.18632/oncotarget.12983

Figure Lengend Snippet: A. Western blot analyzing phosphorylated (p-Akt) and total Akt (t-Akt) after 4h treatment with PTN, PDGFB or the combination of PTN and PDGFB, and quantification of p-Akt normalized to t-Akt (lower panel), Data is shown as mean ± SD from 7 independent experiments. B. Quantification of sphere numbers after stimulation with PTN, PDGFB or PTN in combination with PDGFB. C. - D. Representative image of spheres upon indicated treatment and quantification of sphere size. (mean ± SD; n ≥ 100 spheres/group). E. Immunostaining of olig2, nestin or NG2 in the spheres after the indicated treatment. The experiment was repeated three times. F. Immunostaining of cleaved caspase 3 (cl-casp 3) and phosphorylated histone H3 (P-H3) in spheres after treatment, and quantification of G. cl-casp 3 and H. P-H3 positive cells normalized to total cells (data shown as means ± SD; n ≥ 3 spheres/group). (Bar = 100μm in D, Bar = 20μm in E and F).

Article Snippet: After 2 hours, the cells were treated with 25ng/ml PTN (252-PL-050, R&D systems), 10ng/ml PDGFB (100-14b, Peprotech) or the combination of PTN and PDGFB.

Techniques: Western Blot, Immunostaining

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Development of a reliable automated screening system to identify small molecules and biologics that promote human β-cell regeneration

doi: 10.1152/ajpendo.00515.2015

Figure Lengend Snippet: Function and reported effects of compounds tested on human islet cells

Article Snippet: Function and reported effects of compounds tested on human islet cells table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Compound Stock Concentration Solvent Control Dilution Concentrations Tested Source Catalog No. Dyrk family Harmine 100 mM DMSO 1:100 1, 10 μM Sigma 286044 Neurotransmitters GABA 100 mM PBS 1:20 100, 1,000, 2,500 μM Sigma A2129 Serotonin 10 mM Water 1:40 10, 100, 250 μM Sigma L510041 Adenosine signaling/metabolism NECA 40 mM DMSO 1:100 1, 10, 100 μM Tocaris 1691 UK-432097 1 mM DMSO 1:100 0.1, 1, 10 μM Axon Medchem 1193 A-134974 10 mM Water 1:40 0.1, 1, 10 μM Sigma A2846 Hormones/growth factors Human prolactin 0.1 mg/ml 4 mM HCl/0.1% BSA 1:40 100, 1,000, 2,500 ng/ml R&D Systems 682-PL-050 PDGF 0.025 mg/ml PBS 1:20 50, 500, 1,250 ng/ml Sigma P3076 Erythropoietin 500 U/ml PBS/0.1% BSA 1:20 1, 10, 20 U/ml R&D Systems 287-TC-500 Exendin-4 10 μM PBS 1:20 10 nM California Peptide 507–77 TGF-β superfamily Myostatin 100 μg/ml 4 mM HCl/0.1% BSA 1:40 500, 1,000, 2,500 ng/ml R&D Systems 788-G8-010 Activin A 50 μg/ml PBS/0.1% BSA 1:20 10, 100, 1,000 ng/ml R&D Systems 338-AC-010 Open in a separate window Preparation of compounds tested on human islet cells

Techniques:

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Development of a reliable automated screening system to identify small molecules and biologics that promote human β-cell regeneration

doi: 10.1152/ajpendo.00515.2015

Figure Lengend Snippet: Preparation of compounds tested on human islet cells

Article Snippet: Function and reported effects of compounds tested on human islet cells table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Compound Stock Concentration Solvent Control Dilution Concentrations Tested Source Catalog No. Dyrk family Harmine 100 mM DMSO 1:100 1, 10 μM Sigma 286044 Neurotransmitters GABA 100 mM PBS 1:20 100, 1,000, 2,500 μM Sigma A2129 Serotonin 10 mM Water 1:40 10, 100, 250 μM Sigma L510041 Adenosine signaling/metabolism NECA 40 mM DMSO 1:100 1, 10, 100 μM Tocaris 1691 UK-432097 1 mM DMSO 1:100 0.1, 1, 10 μM Axon Medchem 1193 A-134974 10 mM Water 1:40 0.1, 1, 10 μM Sigma A2846 Hormones/growth factors Human prolactin 0.1 mg/ml 4 mM HCl/0.1% BSA 1:40 100, 1,000, 2,500 ng/ml R&D Systems 682-PL-050 PDGF 0.025 mg/ml PBS 1:20 50, 500, 1,250 ng/ml Sigma P3076 Erythropoietin 500 U/ml PBS/0.1% BSA 1:20 1, 10, 20 U/ml R&D Systems 287-TC-500 Exendin-4 10 μM PBS 1:20 10 nM California Peptide 507–77 TGF-β superfamily Myostatin 100 μg/ml 4 mM HCl/0.1% BSA 1:40 500, 1,000, 2,500 ng/ml R&D Systems 788-G8-010 Activin A 50 μg/ml PBS/0.1% BSA 1:20 10, 100, 1,000 ng/ml R&D Systems 338-AC-010 Open in a separate window Preparation of compounds tested on human islet cells

Techniques: Concentration Assay, Solvent, Control

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Development of a reliable automated screening system to identify small molecules and biologics that promote human β-cell regeneration

doi: 10.1152/ajpendo.00515.2015

Figure Lengend Snippet: Human β-cell proliferation is induced by treatment with some, but not all, compounds tested. A: treatment of human islet cells with harmine at 1 and 10 μM in 5 or 11 mM glucose increased β-cell proliferation. **P < 0.01, 5 mM glucose control vs. 1 μM harmine or 10 μM harmine. *P < 0.01, 5 mM glucose control vs. 1 μM harmine or 10 μM harmine. B and C: neurotransmitters GABA (B) and serotonin (C) had a limited effect on β-cell proliferation, and only treatment with 100 μM GABA at 5 mM glucose was statistically significant *P < 0.05, 5 mM glucose control vs. 100 μM GABA. D–F: compounds involved in adenosine signaling and metabolism [NECA (D), UK-432097 (E), and A-134974 (F)] had a modest effect, with 1 μM UK-432097 and 10 μM A-134974 at 11 mM glucose causing a small but statistically significant increase in β-cell proliferation. *P < 0.05, 11 mM glucose control vs. 1 μM UK-432097 and control vs. 10 μM A-134974. G–J: hormones prolactin (G), PDGF (H), erythropoietin (I), and exendin-4 (J) had no effect on β-cell proliferation other than 1,250 ng/ml PDGF at 5 mM glucose, which caused a small but significant increase in proliferation. *P < 0.05, 5 mM glucose control vs. 1,250 ng/ml PDGF. K and L: treatment with members of the TGF-β superfamily myostatin (K) and activin A (L) had no significant effect on β-cell proliferation. Comparisons between controls or compound treatments at 5 vs. 11 mM glucose were not statistically significant in any compound or control tested, and there was no difference between vehicle controls (P = 0.13–0.62); n = 3–6 donors/treatment condition.

Article Snippet: Function and reported effects of compounds tested on human islet cells table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Compound Stock Concentration Solvent Control Dilution Concentrations Tested Source Catalog No. Dyrk family Harmine 100 mM DMSO 1:100 1, 10 μM Sigma 286044 Neurotransmitters GABA 100 mM PBS 1:20 100, 1,000, 2,500 μM Sigma A2129 Serotonin 10 mM Water 1:40 10, 100, 250 μM Sigma L510041 Adenosine signaling/metabolism NECA 40 mM DMSO 1:100 1, 10, 100 μM Tocaris 1691 UK-432097 1 mM DMSO 1:100 0.1, 1, 10 μM Axon Medchem 1193 A-134974 10 mM Water 1:40 0.1, 1, 10 μM Sigma A2846 Hormones/growth factors Human prolactin 0.1 mg/ml 4 mM HCl/0.1% BSA 1:40 100, 1,000, 2,500 ng/ml R&D Systems 682-PL-050 PDGF 0.025 mg/ml PBS 1:20 50, 500, 1,250 ng/ml Sigma P3076 Erythropoietin 500 U/ml PBS/0.1% BSA 1:20 1, 10, 20 U/ml R&D Systems 287-TC-500 Exendin-4 10 μM PBS 1:20 10 nM California Peptide 507–77 TGF-β superfamily Myostatin 100 μg/ml 4 mM HCl/0.1% BSA 1:40 500, 1,000, 2,500 ng/ml R&D Systems 788-G8-010 Activin A 50 μg/ml PBS/0.1% BSA 1:20 10, 100, 1,000 ng/ml R&D Systems 338-AC-010 Open in a separate window Preparation of compounds tested on human islet cells

Techniques: Control